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Non-coding RNA
In eukaryotic cells, mRNA accounts for about 2% of the genome, and the rest is non-coding RNA. Although non-coding RNA does not have the function of coding proteins, it is widely involved in all aspects of life, such as growth, differentiation, development, immunity, and even plays an important regulatory role in tumor formation. It is one of the research hotspots in the post-genomics era to identify and discover new non-coding RNAs, explore their biological functions and their roles and clinical significance in the occurrence and development of cancer and other diseases.
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  • Review
    YANG Ling, ZHANG Zheng
    Chinese Journal of Biochemistry and Molecular Biology. 2025, 41(11): 1645-1655. https://doi.org/10.13865/j.cnki.cjbmb.2025.08.1152
    Cardiovascular diseases (CVDs) remain one of the leading causes of morbidity and mortality worldwide. In recent years, increasing evidence has highlighted the pivotal roles of circular RNAs (circRNAs) in CVDs. Mitochondrial function, essential for cardiomyocytes to sustain energy metabolism, redox homeostasis, and calcium balance, can be modulated by circRNAs through various mechanisms. These include regulation of mitochondrial DNA (mtDNA) expression and stability, intervention in oxidative phosphorylation (OXPHOS) and reactive oxygen species (ROS) generation, control of mitochondrial calcium homeostasis, and participation in dynamic events such as fusion, fission, and mitophagy. Although the predominant model involves circRNAs acting as “miRNA sponges” in competing endogenous RNA (ceRNA) networks, more direct interactions—potentially with mitochondrial proteins, transcription, or replication factors—have yet to be fully elucidated. From a technical standpoint, current approaches to circRNA identification, functional assays, and mitochondria-specific enrichment remain suboptimal, and the majority of studies rely heavily on cell culture and animal models with limited multi-organ or disease-specific validation. Looking ahead, an integrated multi-omics framework that incorporates spatially resolved transcriptomics, RNA chemical modification profiling and protein interactome analysis will enable a systematic dissection of the circRNA-mitochondrial network, thereby opening new avenues for regulating energy metabolism in cardiomyocytes and vascular cells and for advancing the diagnosis and treatment of cardiovascular diseases.
  • Research Paper
    YANG Yu, DONG Na, ZHANG Chi, HU Zhen-Zhen
    Chinese Journal of Biochemistry and Molecular Biology. 2025, 41(11): 1719-1728. https://doi.org/10.13865/j.cnki.cjbmb.2025.09.1167
    Microglia activation-mediated neuroinflammatory responses serve as a critical pathological basis for the development and progression of various brain diseases. The role of circular RNAs (circRNAs) in the regulation of neuroinflammation is increasingly being recognized. This study aimed to investigate the effect and molecular mechanisms of targeted inhibition of circular RNA Homeodomain Interacting Protein Kinase 3 (HIPK3) (circHIPK3) on lipopolysaccharide (LPS)-induced microglial polarization in BV2 cells. The results showed that LPS stimulation significantly induced polarization of BV2 cells towards the pro-inflammatory M1 phenotype and upregulated circHIPK3 expression (P<0.01). Engineered extracellular vesicles (EVs) with rabies viral glycoprotein (RVG) loaded with circHIPK3 siRNA (RVG-EVs-sicHIPK3) were successfully constructed. Transmission electron microscopy (TEM) revealed their typical EV morphology. nanoparticle tracking analysis (NTA) indicated a peak particle size of 70 nm. And Western blotting analysis confirmed the expression of characteristic membrane marker proteins. Treatment with RVG-EVs-sicHIPK3 significantly suppressed the LPS-induced elevation of inflammatory cytokines (TNF-α, IL-6, IL-1β) in the supernatant and reduced the expression of M1 phenotypic marker proteins (CD16 and CD86) (P<0.01). Concurrently, RVG-EVs-sicHIPK3 increased the number of mitophagosomes within cells, upregulated the ratio of the autophagy-related proteins LC3-II/LC3-I (P<0.01), and downregulated the expression of the autophagy-related protein p62 and mitochondrial-specific proteins (TOMM20 and TIMM23) (P<0.01). The mitophagy inhibitor Mdivi-1 significantly reversed the RVG-EVs-sicHIPK3-mediated downregulation of inflammatory cytokine levels, M1 marker proteins, and mitochondrial protein expression (P<0.01). This study demonstrates that inhibiting circHIPK3 reduces LPS-induced microglial polarization towards the M1 phenotype. The protective mechanism is closely associated with enhanced mitophagic flux and the promotion of damaged mitochondrial clearance.
  • Review
    TIAN Tan, BAO Shan-Jun
    Chinese Journal of Biochemistry and Molecular Biology. 2025, 41(10): 1467-1477. https://doi.org/10.13865/j.cnki.cjbmb.2025.08.1122
    Type 2 diabetes mellitus (T2DM) is a chronic metabolic disorder characterized primarily by pancreatic β-cell dysfunction and insulin resistance. Beyond impaired glucose homeostasis, the glucotoxicity resulting from dysregulated glucose metabolism can lead to complications such as diabetic nephropathy, diabetic coronary heart disease, and diabetic cognitive dysfunction, posing severe threats to patients′ health. Therefore, enhancing glucose metabolism and improving insulin resistance are critical strategies for T2DM prevention and treatment. Currently, the primary therapeutic approaches for T2DM include oral hypoglycemic agents and exogenous insulin injections. However, these methods face challenges such as unclear diagnostic markers, suboptimal therapeutic efficacy, and drug-related side effects. Consequently, the development of more precise and effective treatment strategies remains a top priority in clinical research. MicroRNA (miRNA) is a short non-coding RNA that regulates gene transcription. It has been reported to exhibit significant alterations even before the onset of overt pathological features like hyperglycemia, suggesting their potential as early diagnostic biomarkers for T2DM to address current issues like delayed diagnosis and disease progression due to missed optimal treatment windows. Moreover, miRNAs can modulate the expression of pathways involved in glucose metabolism, thereby improving insulin resistance and maintaining glucose homeostasis while avoiding side effects associated with conventional therapies, such as reduced bone density and localized fat accumulation. This positions miRNAs as a promising avenue for precise and efficient T2DM management. This article systematically elucidates the molecular mechanisms by which miRNAs regulate glucose metabolism-related pathways, providing a theoretical foundation and reference for future clinical research.
  • Research Paper
    LI Yong, QUAN Yi-Ning, WANG Kun, LI Hong-Li, YIN Chong-Gao
    Chinese Journal of Biochemistry and Molecular Biology. 2025, 41(9): 1310-1319. https://doi.org/10.13865/j.cnki.cjbmb.2025.08.1241
    Breast cancer (BRCA) remains one of the leading causes of cancer-related deaths worldwide due to its high rates of metastasis and recurrence, making it crucial to explore its underlying molecular mechanisms. Our previous study demonstrated that miR-326 inhibits BRCA progression by targetingEPH receptor B3(EPHB3). This study further explores the molecular mechanism by which long non-coding RNAs (LncRNAs) regulates BRCA progression via the competing endogenous RNA (ceRNA) mechanism, in which it competes with EPHB3 for miR-326 binding. Bioinformatics analysis identified LncRNA Small Nucleolar RNA Host Gene 12 (SNHG12) as a potential miR-326-binding molecule. SNHG12 was found to be significantly upregulated in BRCA tissues, exhibiting a negative correlation trend with miR-326 and a positive correlation trend with EPHB3, suggesting its potential involvement in the ceRNA regulatory network. Nuclear-cytoplasmic fractionation assays revealed cytoplasmic localization of SNHG12, while dual-luciferase reporter assays confirmed its direct binding to miR-326. Functional experiments demonstrated that SNHG12 knockdown significantly suppressed BRCA cell proliferation, invasion, and migration, while miR-326 inhibition reversed these effects. Furthermore, miRNA pulldown assay revealed significant enrichment of SNHG12 and EPHB3 in the miR-326 pulldown products, indicating direct binding between them. Western blotting and rescue experiments revealed that SNHG12 upregulates EPHB3 expression by sponging miR-326, thereby promoting the malignant behaviors of BRCA cells. Collectively, this study revealed that LncRNA SNHG12 promotes BRCA progression by regulating the miR-326/EPHB3 axis through a ceRNA mechanism. The SNHG12/miR-326/EPHB3 pathway may represent a promising target for the molecular diagnosis and targeted therapy of BRCA.
  • Reviews
    YANG Yi, OUYANG Song-Ying, WU Jun-Jun
    Chinese Journal of Biochemistry and Molecular Biology. 2025, 41(3): 344-352. https://doi.org/10.13865/j.cnki.cjbmb.2025.01.1288
    Circular RNA (circRNA) is a single-stranded RNA with a covalently closed loop structure, which has a more stable structure and lower immunogenicity than linear RNA. Many studies have shown that circRNA has characteristics such as conservation, stability, and tissue specificity, and can act as a microRNA (miRNA) sponge to interact with proteins and translation templates, and regulate biological functions such as gene expression and signal transduction. Based on the characteristics and various biological functions of circRNA, some endogenous circRNA plays an important regulatory role in the occurrence and development of tumors and has the potential to be used as biomarkers and therapeutic targets. In addition, mRNA drugs have limitations such as instability, easy degradation, low translation efficiency, and immunogenicity in practical applications. Engineered translatable exogenous circRNA can solve some of the limitations of linear mRNA application and become a new type of potential efficient drug. In this paper, we introduce the circRNA biogenesis mechanisms, specific biological functions, and the current status of diagnosis and treatment applications in tumors. This includes the diagnostic application of endogenous circRNA in tumors, the design and synthesis strategies of exogenous circRNA, and the current progress in the design and application of engineered circRNA vaccines using their stable and efficient protein expression functions in the treatment of tumors. Finally, we discuss the current clinical diagnostic application problems of circRNA, the challenges of exogenous circRNA therapeutic applications, and the prospects of the field.
  • Reviews
    LI Xin, HU Ying, WANG Yu-Ming
    Chinese Journal of Biochemistry and Molecular Biology. 2025, 41(3): 364-375. https://doi.org/10.13865/j.cnki.cjbmb.2025.01.1194
    The emergence of CRISPR/Cas system has greatly promoted the progress in the field of gene editing, especially CRISPR/Cas9 system, which has become a core tool in biomedical research. Long noncoding RNAs (lncRNAs) play a key role in gene regulation, cell differentiation and the development of a variety of diseases. Especially in cancer research, lncRNAs have important application prospects as cancer biomarkers and therapeutic targets. However, lncRNAs are generally characterized by low abundance and poor conservation, which limits the study of their functions by traditional means. CRISPR/Cas9 technology provides an efficient, flexible and accurate tool for lncRNA research, which significantly accelerates the progress in this field. This paper first reviews the basic principles of CRISPR/Cas9 system and its wide applications in gene editing, including CRISPR knockout, knock-in, interference, activation and other functional systems. These technologies can not only screen key lncRNAs in specific biological processes, but also be used for gene function research to explore their roles in diseases. This article focuses on the analysis of CRISPR/Cas9 technology in the study of lncRNA functions, regulatory mechanisms, and its key applications in tumor research. In addition, the article also summarizes the methods of genome-wide screening by CRISPR/Cas9 to identify functional lncRNAs, and discusses the roles of these lncRNAs in cancer cell proliferation, migration, invasion and drug resistance. CRISPR/Cas9 knockout system can efficiently knock down lncRNA genes and reveal their specific functions in gene regulation. At the same time, CRISPR activation and interference technology provide a new idea for the research of noncoding genes, and further explore its clinical application in cancer and other diseases by regulating the expression level of lncRNAs. The article also discusses the potential of CRISPR technology in future lncRNA research, especially the progress in solving technical problems such as genome complexity, targeting efficiency and off-target effects. As mentioned in the review, CRISPR/Cas9 technology not only provides a powerful tool for studying lncRNAs, but also provides new ideas and opportunities for developing new means of cancer diagnosis and treatment in the future.
  • Reviews
    YANG Xin-Ling, ZHANG Dong-Dong, CHANG Xiao-Tong
    Chinese Journal of Biochemistry and Molecular Biology. 2025, 41(3): 384-392. https://doi.org/10.13865/j.cnki.cjbmb.2024.12.1299
    Liquid-liquid phase separation (LLPS) is the process where intracellular biomolecules rapidly form reversible high concentrations of liquid-phase condensates, resulting in the compartmentalization and the formation of intracellular membraneless organelles. LLPS is involved in various biological processes and pathologic processes, such as neurodegenerative diseases and cancers. Long noncoding RNAs (lncRNAs) have been revealed to be closely related to phase separation. It has become a research hotspot in life science recently, because it provides new insight into the mechanism of lncRNAs. In this review, we focuses on the effect of lncRNA SLERT on the phase separation of nucleolar fibrillar center/dense fibrillar component (FC/DFC) by interacting with DDX21 protein as a molecular chaperone; LINC00657 (NORAD) forms NP bodies with PUM proteins, which drives PUM protein liquid droplets and inhibits its activity, and promotes genomic stability; DilncRNA regulates DNA damage response small RNAs (DDRNAs) and LLPS of p53 binding protein 1 (53BP1) in response to DNA damage, and lncRNA LINP1 phase separation droplets bind to Ku protein to promote DNA damage repair; LncRNA SNHG9, MELTF-AS1 and MALR drive LATS1, YBX1 and ILF3 protein LLPS respectively to promote cancer, while GIRGL and lncFASA act as tumor suppressor genes in cancer development through regulating the phase separation of CAPRIN1 and PRDX1 respectively; LncRNA XIST drives X chromosome inactivation by LLPS. In a word, we summarize the latest research progress about the functional roles of lncRNAs in FC/DFC nucleolus, genomic stability and DNA damage and repair, cancer and X-chromosome inactivation through regulating LLPS. This paper shows that lncRNAs can participate in multiple pathophysiological processes by regulating LLPS, which is expected to provide a new direction for the treatment of LLPS-mediated diseases.
  • Research Papers
    LIU Yi-Lan, HU Ya-Mei, LI Hui-Xia, LU Jia-Wei, LIU Yuan
    Chinese Journal of Biochemistry and Molecular Biology. 2025, 41(2): 284-295. https://doi.org/10.13865/j.cnki.cjbmb.2024.12.1268
    Yanchi Tan sheep, a specialty of Ningxia, China, is famous for its excellent meat quality, but the detailed mechanisms of its muscle development and meat quality formation are still unclear. In order to preliminarily investigate the unique molecular mechanisms in the muscle development of the sheep, the present study was carried out to collect the longest dorsal muscle (LDM) samples from 6-month-old and 12-month-old female Tan sheep, and we screened the genes and long non-coding RNA(lncRNAs) that may play important roles in regulating the muscle development and meat quality formation through transcriptome sequencing(RNA-seq). The results showed that a total of 504 mRNAs and 1,483 lncRNAs were detected between the two groups of LDMs, with significant differences (P<0.05). And we also predicted 103 potential lncRNA-mRNA cis-regulatory relationship pairs with significant differences (P<0.05). Skeletal muscle development-related genes such as MSTN, CDKN1A, FHL1 and skeletal muscle metabolism-related genes such as LPL and SCD were subsequently screened based on the functional analysis of differentially expressed mRNA. Based on the functional analysis of differentially expressed lncRNA target genes, 13 lncRNAs potentially related to muscle development (LNC.17926.1, LNC.981.1, LNC.16284.1, etc.) and 8 lncRNAs potentially related to skeletal muscle metabolism (LNC.15496.1, LNC.18149.1, etc.) were identified. 5 lncRNAs and 4 mRNAs expressed in both groups were randomly selected for qPCR validation experiments, and their results were consistent with RNA-seq, which corroborated with the reliability of the sequencing results. This study lays a solid foundation for future investigations of muscle development, meat quality formation mechanism and molecular breeding in Tan sheep, and provides a scientific basis for improving the meat production performance of other domestic sheep breeds.
  • Reviews
    ZHU Chun-Yuan, LI Fei, WANG Yu-Ping
    Chinese Journal of Biochemistry and Molecular Biology. 2025, 41(1): 89-98. https://doi.org/10.13865/j.cnki.cjbmb.2024.10.1315
    Alpha-enolase 1 (ENO1) is a multifunctional protein that serves as a glycolysis enzyme, a plasminogen receptor, and a DNA-binding protein, playing a critical role in tumorigenesis and progression. Long non-coding RNAs (lncRNAs) are crucial regulators of chromatin structure and gene expression. Accumulated data have unveiled that lncRNAs regulate ENO1 through multiple pathways. In the nucleus, lncRNAs act on the ENO1 promoter to regulate its transcription. In the cytoplasm, lncRNAs affect ENO1 translation by acting directly or indirectly with ENO1 mRNA and by adsorbing small ribonucleic acids as sponges. Additionally, in the cytoplasm, lncRNAs influence the stability, enzyme activity, and other functions of ENO1 by directly binding to the ENO1 protein and acting as scaffold molecules to form a polymer with ENO1. In tumors, the abnormal expression of lncRNA regulates the overexpression and function of ENO1, promoting the proliferation and metastasis of tumor cells. There is currently some knowledge on the interaction between lncRNA and ENO1, and further investigation is needed to reveal the specific molecular mechanisms and biological functions, as well as the potential applications of lncRNA in clinical treatment. Here, we review the regulatory role of lncRNAs on ENO1, focusing on 16 identified lncRNAs that regulate ENO1. Understanding the regulatory effects of lncRNAs on ENO1 will enhance our knowledge of the regulatory network involving ENO1 in tumor cells and may reveal new targets for cancer treatment.
  • Research Papers
    YU Ya-Nan, LI Jia-Qiu, MA Xiao-Lin
    Chinese Journal of Biochemistry and Molecular Biology. 2025, 41(1): 156-168. https://doi.org/10.13865/j.cnki.cjbmb.2024.12.1242
    The long non-coding RNA KCNQ1OT1 plays an important role in promoting the occurrence and development of various cancers. However, there is currently no systematic analysis of KCNQ1OT1 in pan cancer. To elucidate the value of KCNQ1OT1 in tumor diagnosis and prognosis, this study analyzed its expression levels in pan-cancer tissues and its impact on patient prognosis. By analyzing the regulatory mechanism of KCNQ1OT1 in gastric cancer, new molecular targets may be found for the diagnosis and treatment of gastric cancer. Using Sangerbox 3.0, ACLBI and UALCAN databases, we found the expression levels of KCNQ1OT1 were increased in 7 tumor tissues types (P<0.05). We found KCNQ1OT1 expression was correlated with poor prognosis in many tumor types using Sangerbox 3.0 database. We used R software to analyze the differential genes between the high and low expression groups of KCNQ1OT1 in gastric cancer patients (P<0.05, log2FoldChange>1). The GO and KEGG enrichment analysis showed that KCNQ1OT1 was involved in the glutamine metabolism of gastric cancer. The cell counting and Western blot detection showed that knocking down KCNQ1OT1 significantly reduced the gastric cancer cell activity, SLC1A5 expression level and SLC1A5-mediated glutamine transport process (P<0.01). Bioinformatics, RNA immunoprecipitation and dual luciferase analysis confirmed that KCNQ1OT1 competitively bind to miR-138-5p to promote the expression of SLC1A5. Finally, ChIP-seq data was used to detect the high H3K27ac signaling at the gene locus of KCNQ1OT1, and ChIP-qPCR was used to verify that P300-mediated enhancer activity regulated the high expression of KCNQ1OT1 in gastric cancer. KCNQ1OT1 can serve as an independent diagnostic biomarker and prognostic predictor in various tumors. Targeting the KCNQ1OT1/miR-138-5p/SLC1A5 signaling axis to regulate glutamine metabolism may provide new strategies and molecular targets for the treatment of gastric cancer.
  • Reviews
    ZHANG Yi-Xin, WANG Xiang, LU Yu-Zhu
    Chinese Journal of Biochemistry and Molecular Biology. 2024, 40(12): 1691-1697. https://doi.org/10.13865/j.cnki.cjbmb.2024.10.1118
    microRNA is a class of important, regulatory and small RNA in eukaryotes. Plant microRNAs play key roles in regulating growth and development as well as responses to the environment. Although some reviews about plant microRNA have been published, few of them address how microRNAs regulate agronomic traits of crops. Here we review how microRNA can affect the expression levels of various enzymes, functional proteins and transcription factors, including laccase, antioxidant enzymes, DELLA proteins, stress tolerance proteins, auxin response factors, proliferative cytokines, etc., and also regulate important bioactive substances such as lignin, erythromycin, proline and glutathione in crops. Through distinct mechanisms, microRNAs modulate material transport and metabolism, organ development, and stress resistance of crops under various stresses, and play an important role in the crop morphology, biomass, grain yield, nutrient metabolism, and stress response, etc. Finally, we discuss the potential of these microRNAs in agricultural production, and propose feasible strategies and directions for future crop improvement and breeding.
  • Research Papers
    FU Ya-Kun, JIA Lin-Chuang, MU Biao
    Chinese Journal of Biochemistry and Molecular Biology. 2024, 40(12): 1698-1708. https://doi.org/10.13865/j.cnki.cjbmb2024.11.1307
    Colorectal cancer is one of the most common malignant tumors of the digestive system. Oxaliplatin(OXA)-based combination chemotherapy is the most commonly used strategy for treating patients with advanced-stage disease in clinical practice. However, the development of resistance greatly limits the effectiveness of chemotherapy and is a major cause of treatment failure. Due to the unknown mechanisms of resistance, there is an urgent need for a high-throughput, highly specific sequencing method to explore the causes of oxaliplatin resistance. Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated protein 9 (CRISPR/Cas9) is a rapidly advancing high-throughput technology that can be employed for screening resistance genes. However, its role in identifying genes involved in oxaliplatin resistance in colorectal cancer remains unclear. We constructed an sgRNA library containing 5 256 small-guide RNAs (sgRNAs) targeting 910 human epigenetic-related genes, using lentivirus packaging. By determining the viral infection conditions through protein immunoblotting and flow cytometry, we maintained the multiplicity of infection (MOI) below 30% to ensure that each cell is infected with only one sgRNA, thereby knocking out one gene. Colorectal cancer cells HCT116 and SW620 were infected with lentivirus carrying library, and single clones were obtained and expanded through a positive selection strategy. By the positive selection strategy, we identified 21 genes that regulate the sensitivity of colorectal cancer cells to oxaliplatin. By knocking out of candidate genes, we observed that deletion of TDRKH, ALKBH3, UNKL, TTF2, TNKS, AURKA, RBM12, ELAVL2, DKC1, LSM5, NOL8 and PRPF3 significantly increased the half-maximal inhibitory concentration (IC50) of oxaliplatin in colorectal cancer cells (P<0.05). Among them, high expression of the ALKBH3, AURKA, and RBM12 genes was significantly correlated with clinical prognosis (overall survival: P=0.043, P<0.0001, P=0.045; recurrence-free survival: P=0.004, P=0.0019, P=0.0064). Our study demonstrates that the CRISPR/Cas9 library is a high-throughput method for screening tumor sensitivity genes, providing target references for further exploring the mechanism of colorectal cancer sensitivity to oxaliplatin.
  • Reviews
    WANG Shu-Ji, CAO Ru-Fei, DUAN Xiao-Lei
    Chinese Journal of Biochemistry and Molecular Biology. 2024, 40(7): 889-896. https://doi.org/10.13865/j.cnki.cjbmb.2024.01.1419
    MicroRNAs (miRNAs) are small non-coding RNAs that are closely associated with the occurrence and progression of tumors and other diseases. However, miRNAs require high-sensitivity and high-specificity detection due to their low abundance, high sequence homology, and rapid degradation. The RNA-cleaving deoxyribozyme (RCD) is a functional single-stranded DNA molecule that enables specific cleavage of substrates to release miRNAs that can be recycled with the assistance of metal ions, prompting cyclic signal amplification. Recently, developing new methods based on RCD catalytic amplification for miRNA high-sensitivity detection has become the focus of researchers. Based on combing with the various new technologies and materials in miRNA biosensors, this study classifies and reviews the new methods for detecting miRNAs based on deoxyribozyme catalytic amplification developed in recent years. We separate these miRNA detection strategies into three categories: RCDs combined with DNA self-assembly, isothermal amplification, and nanomaterials. We explore the basic principles of each approach, the latest research advancements, and application scenarios in biomedical sensors and medical detection. This review provides a foundation and reference for further research of highly sensitive and accurate miRNA detection strategies.
  • Research Papers
    WU Hai-Li, DUAN Bing, DU Jin-E, MA Jia-Jing, LI Zhuo-Yu
    Chinese Journal of Biochemistry and Molecular Biology. 2024, 40(7): 964-975. https://doi.org/10.13865/j.cnki.cjbmb.2024.05.1459
    In recent years, the application of long non-coding RNAs (lncRNAs) in the treatment of diseases has been widely concerned. LncRNAs have been proved to play a crucial role in tumor therapy. They usually act as oncogene or suppressor genes to regulate the occurrence and development of tumors. Previously, our group discovered a new lncRNA 606938 that can be used as a target in colorectal cancer through transcriptome analysis. However, its molecular mechanism in colorectal cancer is still unclear. In this study, RT-qPCR test shows that lncRNA 606938 is low expressed in colorectal cancer cell lines compared with normal colon epithelial cells. The clonogenicity was decreased by 97% and 54.5% in lncRNA 606938 overexpressed DLD-1 and SW620 cells, respectively. The results of flow cytometry assay showed that overexpression of lncRNA 606938 caused cell cycle arrest at G1 phase (They were prolonged by 50.5% and 63.9%, respectively) and promoted apoptosis of colorectal cancer cells, which was increased by 1.9% and 3.3%, respectively. Western blot results showed that overexpression of lncRNA 606938 down-regulated the expression of related oncogenes (β-catenin, c-Myc, cyclin D1, cyclin D2, cyclin D3, CDK 4, CDK 6) in colorectal cancer cells, and the expression of tumor suppressor genes (TRIM33, caspase 2, and actived caspase 3) was up-regulated. The above results were reversed after knockdown of lncRNA 606938. Mechanistically, it has been confirmed through RNA pulldown assay, RIP assay, and Rescue assay that lncRNA 606938 can target and promote TRIM33 expression, thereby promoting the degradation of β-catenin and blocking the expression of β-catenin and its downstream oncogenes such as c-Myc and cycline D1 to inhibit the progression of colorectal cancer. This study elucidates the molecular mechanism by which the lncRNA 606938 targets TRIM33/ β-catenin signaling pathway, inhibits the proliferation and promotes the apoptosis of colorectal cancer cells. This study reveals the potential of lncRNA 606938 as a tumor suppressor gene, providing new target and strategies for the clinical treatment of colorectal cancer.
  • Research Papers
    WU Hao, ZHANG Wen-Tao, JIA Feng-Feng, LIU Ming, ZHU Jian-Jun
    Chinese Journal of Biochemistry and Molecular Biology. 2024, 40(7): 976-986. https://doi.org/10.13865/j.cnki.cjbmb.2024.05.1037
    Long non-coding RNA (lncRNA) PART1, a competing endogenous RNA (ceRNA), plays a crucial role in the occurrence and development of various tumors. However, research on PART1 in laryngeal squamous cell carcinoma (LSCC) remains scarce. Based on preliminary lncRNA sequencing data, we found that PART1 was significantly downregulated in LSCC tissues. Further analysis of sequencing and clinical data from public databases such as TCGA revealed 146 differentially expressed lncRNAs (95 upregulated and 51 downregulated) and 2 424 differentially expressed mRNAs when comparing LSCC tumor and adjacent tissues. The results showed that PART1 was generally downregulated in LSCC (P < 0.0001), and patients with high PART1 expression had significantly better prognosis (P < 0.05). We used bioinformatics methods to construct the ceRNA regulatory network of PART1 in LSCC and identified the miRNAs and mRNAs interacting with it. Under laboratory conditions, the importance of PART1 in LSCC cells was validated in vitro. Overexpression vectors significantly increased the expression of PART1 in LSCC cells (P < 0.001). Experiments including 5-ethynyl-2′-deoxyuridine staining, apoptosis analysis, scratch healing assay, Transwell assay, and phalloidin staining showed that overexpression of PART1 significantly affected the proliferation, apoptosis, migration, and invasion of LSCC cells in vitro (P < 0.001). Therefore, PART1 may suppress the occurrence and development of LSCC. This study provides a theoretical basis for elucidating the role of PART1 in LSCC.
  • Research Papers
    LI Yi, WEN Yi-Hong, WU Hua-Yan, JIANG Jia-Xue, OU Tao, CHEN Kai-Yin, LIU Yu-Peng, SHAN Zhi-Xin
    Chinese Journal of Biochemistry and Molecular Biology. 2024, 40(4): 544-553. https://doi.org/10.13865/j.cnki.cjbmb.2024.02.1002
    The increasing evidence suggests that long noncoding RNAs play an important role in regulating biological functions. Comparing with the healthy organ donors (n=23), the results of real-time quantitative polymerase chain reaction (RT-qPCR) assay showed that long noncoding RNA RP11-879F14.2 was significantly increased in the myocardium of patients with heart failure (HF) (n=21), however, the role and mechanism of RP11-879F14.2 in cardiac hypertrophy remains unclear. The effect of adenovirus-mediated overexpression of RP11-879F14.2 on the expression of hypertrophy-related genes, including myosin heavy chain 7 (MYH7),skeletal muscle actin alpha 1 (ACTA1) and natriuretic peptide type A (NPPA), was evaluated, and the RT-qPCR results revealed that overexpression of RP11-879F14.2 could markedly inhibit the expression of cardiac hypertrophy-related genes in neonatal mouse ventricular cardiomyocytes (NMVCs) and neonatal rat ventricular cardiomyocytes (NRVCs). The results of RT-qPCR and Western blotting showed that RP11-879F14.2 could efficiently enhance the expression of pyruvate kinase M2 (PKM2) in NMVCs. Overexpression of PKM2 and RP11-879F14.2 could consistently attenuate the hypertrophy-related genes expression in NMVCs and NRVCs, and inhibited the increase of cell size of phenylephrine (PE)-induced NRVCs. Moreover, knock-down of PKM2 could reverse the inhibitory effect of RP11-879F14.2 on the cardiac hypertrophy-related genes expression in NMVCs. The glucose metabolic alterations were accessed by using Seahorse XF96 extracellular flux analyzer. Overexpression of RP11-879F14.2 and PKM2 could consistently enhance glucose metabolism in NMVCs, but knock-down of PKM2 could inhibit some RP11-879F14.2-promoted glycolysis-related genes, TCA cycle-related genes and mitochondrial ETC-related genes expression in NMVCs. Therefore, RP11-879F14.2 inhibits cardiomyocyte hypertrophy via upregulating PKM2 expression.
  • Reviews
    WANG Qin, HE Meng-Jie, LONG Fang-Yi
    Chinese Journal of Biochemistry and Molecular Biology. 2024, 40(3): 295-311. https://doi.org/10.13865/j.cnki.cjbmb.2024.01.1343
    Circular RNA (circRNA) is a special class of non-coding RNA (ncRNA) in eukaryotic cells, which is highly stable. At present, circRNA has been widely found to be involved in the occurrence and development of various diseases as microRNA (miRNA) sponges. As a research hotspot in recent years, the function and effects of circRNA are constantly being discovered and utilized. CircRNAs can act as protein scaffolds to bind targets more tightly and regulate transcription of disease signaling molecules, as well as participate in protein expression as translation templates. A substantial body of research has established the pivotal role of ferroptosis in disease development. Iron and lipid metabolism-related targets are recognized as key components in the regulation of ferroptosis. Recent studies have revealed that circRNA not only participates in ferroptosis regulation as a miRNA and protein scaffold but also exerts control over ferroptosis-related signaling molecules at the transcriptional level. Currently, synthetic circRNA molecules have been engineered for the development of RNA vaccines. Additionally, a range of gene therapy technologies and drug delivery vectors offer a promising foundation for the development of therapeutics targeting circRNA. This paper describes the functions and effects of circRNA, its role as a key target in the regulation of ferroptosis, and its implications in various diseases. Special emphasis is placed on discussing the role of circRNA in multisystem diseases, including tumors, nervous system disorders, and diabetes-related complications. Additionally, drug development strategies related to circRNA are summarized to provide insights for the development of therapeutic drugs targeting ferroptosis.
  • Reviews
    CHEN Yi-Yu, YE Ting
    Chinese Journal of Biochemistry and Molecular Biology. 2024, 40(2): 214-222. https://doi.org/10.13865/j.cnki.cjbmb.2023.08.1091
    Circular RNAs (circRNAs), which are close-looped and high stabile RNA molecules, have gradually attracted scientific interest for their biological function in the physiological and pathological process, especially in cancer.Cancer stem cells (CSCs), which are a subpopulation of tumor cells that possess the characteristics of self-renewal, aberrant differentiation potential, tumorigenesis and therapeutic resistance. The theory of CSCs opens a new dimension for tumor cell biology and clinical oncology research, which needs in-depth studies by researchers and clinicians. In this review, we cover biological features of CSCs, the biological structure, function and its detection technology of circRNAs, the regulatory mechanism of circRNAs in CSC phenotypes such as self-renewal, differentiation, proliferation and invasion. Furthermore, we explore the potential clinical value of circRNAs as CSC biomarkers for clinical diagnosis, treatment and prognosis.
  • Research Papers
    WU Yuan, LI Zu-Ming, WU Si-Yi, CHEN Jian-Kun, LI Ji-Qiang, CHEN Hai, CAI Shu-Bin
    Chinese Journal of Biochemistry and Molecular Biology. 2023, 39(11): 1576-1587. https://doi.org/10.13865/j.cnki.cjbmb.2023.09.1185
    Intermittent hypoxia (IH) is an important pathophysiological feature of obstructive sleep apnea (OSA), but its molecular mechanism is still unclear. We aim to investigate the role of endogenous competing endogenous RNA (ceRNA) regulatory network in the development of IH in OSA rats. An intermittent hypoxic rat model of OSA was constructed by hypoxic and reoxygenation cycles. CircRNAs and mRNAs were detected in rat bronchial tissues, and 230 up-regulated and 181 down-regulated circRNAs and 1238 up-regulated and 608 down-regulated mRNAs were analyzed and screened. The results of Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of the differential circRNAs and mRNAs suggested that they were mainly associated with metabolic pathways and PI3K-Akt signaling pathways. The key circRNAs (the top six circRNAs with the largest differences) were further validated by quantitative real-time polymerase chain reaction (qRT-PCR), chr9:52042693|52047844 and chr4: 64889575|64899587 were expressed in bronchial tissues consistent with the sequencing results, which were used to further construct the ceRNA regulatory network. Four potential ceRNA regulatory networks were identified by TargetScan and miRanda database, combined with the results of differential circRNA and mRNA. The expression of molecules in the four potential ceRNA regulatory networks was detected by qRT-PCR in bronchial and lung tissues, and the results suggested that the expression of this regulatory network, chr9:52042693|52047844-miR-351-5p-Pten, was consistent with the sequencing results. The findings indicate that chr9:52042693|52047844-miR-351-5p-Pten may be involved in the development and progression of obstructive sleep apnea syndrome through a ceRNA mechanism.
  • Lipid Metabolism Column
    DANG Yuan-Yuan, WANG Lei, JIA Lin-Tao, ZHANG Rui, WANG Xiao-Jian
    Chinese Journal of Biochemistry and Molecular Biol. 2023, 39(5): 663-671. https://doi.org/10.13865/j.cnki.cjbmb.2023.03.1405
    The long non-coding RNA CASC15 (CASC15) is considered as a tumor-related lncRNA involved in regulating biological processes such as proliferation, invasion and migration of a variety of tumor cells. However, the role of CASC15 in intracellular lipid metabolism remains unclear. The sterol-regulatory element binding proteins 1 (SREBP1) is a key transcription factor that regulates lipid metabolism in cells, and its member SREBP1a mainly regulates the expression of key enzyme genes in lipid synthesis. In this paper, we investigated the effects of CASC15 on the expression and localization of lipid regulator SREBP1a in human liver cells through molecular biology and cell function experiments. The results showed that in liver cells, the mRNA and total protein levels of intracellular SREBP1a remained unchanged after overexpression of CASC15 (P<0.001), while the precursor protein levels increased (P<0.05) with enhanced translocation into the nucleus; However, the levels of fatty acid synthesis-related products free fatty acids (P<0.001) and triglycerides (P<0.001) regulated by SREBP1a showed a down-regulation trend. This paper reveals a possible mechanism by which CASC15 regulates lipid metabolism in liver cells, hoping to provide new ideas for the treatment and research of diseases related to lipid metabolism disorders.
  • Reviews
    ZHANG Ya-Ting, LI Wen-Na, PENG Wei-Pan
    Chinese Journal of Biochemistry and Molecular Biol. 2023, 39(5): 692-698. https://doi.org/10.13865/j.cnki.cjbmb.2022.09.1226
    MicroRNAs (miRNA) are non-coding RNAs with short sequences that play important regulatory roles in biological process such as cell proliferation, differentiation and apoptosis. The occurrence and development of many diseases are related to abnormal expression of miRNA. Therefore, accurate detection of relevant miRNA at the early stage of disease occurrence will contribute to early diagnosis and timely treatment of diseases. Nucleic acid biosensors have been widely applied in detection due to stable chemical properties, simple design, easy modification and programmability. The G-quadruplex functional nucleic acids are special spatial structure formed by guanine-rich DNA through Hoogsteen hydrogen bonds. The G-quadruplex has been widely used in miRNA detection because it can interact with some small molecules such as porphyrin, fluorescein and hemin to enhance their physicochemical properties through Van der Waals forces andhydrophobic interaction and other interactions. At present, the G-quadruplex has been combined with nucleic acid amplification or nano-signal amplification technology to improve sensitivity, which can be used for target recognition and auxiliary signal output. Firstly, this paper introduced the structure of G-quadruplex and the mechanism to enhance fluorescence and increase enzyme-like activity. Secondly, according to the above characteristics, the research progress of G-quadruplex combined with fluorescent ligands to construct miRNA fluorescent biosensors, and the miRNA biosensing platform based on G-quadruplex/hemin in colorimetric, electrochemistry and chemiluminescence detection were discussed. Finally, the challenges faced by G-quadruplexes in miRNA detection were discussed, and the future development trend was prospected. This review may help to promote the construction of nucleic acid biosensors with high sensitivity and specificity.
  • Research Paper
    WANG Chen-Yan, DU Jiang, MA Zhen-Nan, ZHANG Li, WANG De-Fu, NIU Yan-Bing
    Chinese Journal of Biochemistry and Molecular Biol. 2023, 39(5): 724-733. https://doi.org/10.13865/j.cnki.cjbmb.2023.03.1617
    MicroRNAs (miRNAs) are small non-coding RNAs that play an important role in regulating target genes in stress response and biotic stress in plants. It is unclear whether endogenous miR167 of Pinellia ternate is involved in virus stress. In this study, the endogenous miR167 of Pinellia ternate was firstly amplified and named pt-miR167. The conservative analysis, phylogenetic analysis and target gene prediction were carried out using bioinformatics software. On this basis, qRT-PCR was used to detect the accumulation of soybean mosaic virus(SMV), the expression level of miR167 and its potential target genes, analysis of miR167 and target gene expression patterns in response to viral infection. The results showed that the virus accumulation increased rapidly at 5~15 d, with the fastest increase at 10 d, and then showed a slow upward trend. After virus infection, compared with the control group, the expression of pt-miR167 was down-regulated, and the expression was the lowest at 10 d. Evolutionary tree analysis shows that, pt-miR167 is highly homologous to sly-miR167a in tomato (Solanum lycopersicum). The predicted expression of ARF6 target genes is in the opposite trend to that of pt-miR167, and its expression reached the highest level at 10 d. This study shows that miRNA participates in the interaction between the host and the virus. The results are helpful to reveal the interaction mechanism between SMV and the host Pinellia ternate. It will provide a reference for further studies of viruses and plants in the future, and a research basis for the production of Pinellia ternate.
  • Reviews
    WANG Zhen, ZHANG Le, ZHANG Yu-Zhe
    Chinese Journal of Biochemistry and Molecular Biol. 2023, 39(2): 222-233. https://doi.org/10.13865/j.cnki.cjbmb.2022.06.1082
    Circular RNA (circRNA) are a class of single-stranded covalently closed RNA molecules formed by back-splicing. It can regulate various life activities through sponge microRNA(miRNA), bind to RNA-binding protein (RBP), and regulate gene expression. In addition, circRNA can also perform translation activities and is considered to be a promising biomarker. N6-methyladenosine(m6A) is the most widespread and most common way of RNA modification in eukaryotes. It functions through three types of regulators: m6A methyltransferases (writers), m6A demethylases (erasers) and m6A recognition proteins (readers). In addition to its role in mRNA, m6A also has a regulatory role in circRNA. m6A modification can regulate the expression, stability, cytoplasmic transfer, translation, and evasion of innate immunity of circRNA, and it has been found to be involved in colorectal cancer, liver cancer, non-small cell lung cancer, cervical cancer, breast cancer, osteosarcoma, hypopharyngeal squamous cell carcinoma, pancreatic ductal adenocarcinoma, gastric cancer and other tumors. In addition, m6A modifications have been found to regulate immune responses. In this paper, the regulatory mechanism of m6A modification based on circRNA was reviewed, the regulatory role of m6A modified circRNA in various tumors and immune responses was expounded, and the effect of m6A modified circRNA on immune response was discussed. m6A modification may enhance circRNAs as biomarkers in multiple ways, and put forward a new perspective for the diagnosis, treatment and prognosis of diseases based on m6A modification of circRNA in the future.
  • Research Papers
    XU Hua, ABULIZI Maierha-Ba, ZHAO Wuan-Zhu, HE Li-Juan
    Chinese Journal of Biochemistry and Molecular Biol. 2023, 39(2): 285-294. https://doi.org/10.13865/j.cnki.cjbmb.2022.12.1652
    Recent studies have shown that long-term heavy smoking can cause irreversible damage to the testis and then block spermatogenesis, in which the apoptotic signaling pathway plays a key role. However, its molecular regulation mechanism remains to be further studied. The purpose of this study is to explore the targeting regulation mechanism of miRNA-138-5p in the injury of testicular Sertoli cells (TM4) induced by cigarette smoking. The MTT assay, lactate dehydrogenase (LDH) assay and TUNEL assay showed that after cigarette smoke exposure (CSE) intervention, the survival rate of TM4 cells was significantly decreased (P<0.05), LDH activity was significantly increased (P<0.05), and apoptosis rate was significantly increased (P<0.05). RT-PCR and Western blotting results showed that the mRNA and protein expression levels of p53, Bak and caspase-3 were significantly up-regulated after 10% CSE intervention (P<0.05). CSE intervention was also performed after transfection of miRNA-138-5p overexpression plasmids. The results showed that the survival rate of Sertoli cells was significantly increased (P<0.05), the number of apoptotic positive cells was significantly decreased (P<0.05), and the mRNA and protein expressions of p53, Bak and caspase-3 were significantly down-regulated compared with those before transfection (P<0.05). After silencing miRNA-138-5p, the mRNA and protein expression levels of Bak and Caspase-3 were significantly increased (P < 0.05). Online database analysis showed that miRNA-138-5p and caspase-3 had high matching prediction values. The double luciferase reporter gene results showed that Bak did not bind to miRNA-138-5p, while caspase-3 could bind to miRNA-138-5p. These results suggest that miRNA-138-5p can target the regulation of Caspase-3 and slow down the apoptosis of Sertoli cells induced by cigarette smoking, which has a protective effect on Sertoli cells.
  • Research Papers
    DUAN Yi-Fan, GU Jing, SHU Ya-Fei, HAN Xiao-Fei, LIANG Qian-Kun
    Chinese Journal of Biochemistry and Molecular Biol. 2023, 39(1): 108-120. https://doi.org/10.13865/j.cnki.cjbmb.2022.11.1330
    In recent years, it has been found that microRNA (miRNA) carried by exosomes play an important role in the development of tissue and organ fibrosis. However, studies on the relationship between radiation-induced cardiac fibrosis (RICF) and exosomes and their miRNA are very limited. The purpose of this study is to analyze the possible role of radiation-induced exosome miRNA in the development of RICF by bioinformatics. Myocardial fibroblasts (CFs) are the main effector cells of RICF. The CFs were irradiated with 2 Gy X-rays, and radiation-induced exosomes (X-exo) and unirradiated exosomes of CFs (Exo) were extracted by overspeed centrifugation. Then, exosomes were observed and identified with its morphology, NTA concentration, and exosome surface marker proteins such as CD9, CD63, and CD81. Subsequently, RNA-seq technology was used to detect the miRNA expression profiles of Exo and X-exo, then the differentially expressed miRNAs were screened and their potential target genes and enrichment analysis were analyzed. The results showed that, compared with the control group (Exo), 9 miRNAs were up-regulated in the X-exo (|log2 Fold Change|>1, P<0.05), of these, 8 are the ones with |log2 Fold Change|>2; 19 miRNAs were down-regulated in the X-exo (|log2 Fold Change|>1, P<0.05), of these, 12 are the ones with |log2 Fold Change|>2. TargetScan, miRWalk and miRNADB were used to predict the target genes of differentially expressed miRNAs, and GO and KEGG enrichment analysis were performed. GO enrichment results showed that target genes regulated by differentially expressed miRNAs were mainly involved in biological processes such as protein phosphorylation and cell signal transduction. Those differentially expressed miRNAs were enriched in cytoplasm, cell membrane and other cellular components, playing molecular functions such as protein kinase binding and protein binding. KEGG enrichment results showed that target genes with differentially expressed miRNAs were mainly enriched in PI3K-Akt, MAPK, mTOR, ECM-receptor interaction, cAMP, Wnt, TGF-β and Notch related signaling pathways. Combing with literatures, it is suggested that differentially expressed exosomal miRNA may promote the development of RICF by regulating the target genes and related signaling pathways. Because the activation regulation of signaling pathways such as MAPK, PI3K/AKT, and mTOR are mainly dependent on the phosphorylation of key molecules, but not on the regulation of the transcriptional level. The detailed mechanism of "Exosomes miRNA mediate radiation bystander effects to promote RICF" will be investigated in future studies, and the signaling pathways such as ECM, cAMP, TGF-β, Wnt, Notch, Ras, and Rap1 should be the focus.
  • Research Papers
    KUANG Ye, DONG Yu-Lin, JI Yong, CAO Xiang-Hong
    Chinese Journal of Biochemistry and Molecular Biol. 2023, 39(1): 130-141. https://doi.org/10.13865/j.cnki.cjbmb.2022.12.1414
    Hypertrophic cardiomyopathy (HCM) is a kind of primary cardiomyopathy characterized by hypertrophic cardiomyocytes. The molecular mechanism of its pathogenesis has not been clearly defined. This study integrates the data sets numbered GSE130036 and GSE36946 in the Gene Expression Omnibus (GEO) database. The expression difference analysis of RNA expression matrix in the data set are performed by R language. The biological function enrichment analysis of differentially expressed mRNAs was carried out, and the chromatin remodeling factor BPTF (bromodomain PHD finger transcription factor) was identified. The binding sites of miRNA targeting mRNA and lncRNA were predicted with BPTF as the core, and the endogenous competitive RNA (ceRNA) network of lncRNA TNRC6CAS1-miR-30c-1-3p-BPTF was constructed. The expression of BPTF and lncRNA TNRC6CAS1 was significantly up-regulated (P<0.001), while the expression of miR-30c-1-3p was significantly down-regulated (P<0.001) in HCM clinical samples (peripheral blood of 20 healthy people and 20 hypertrophic cardiomyopathy patients) and myocardial hypertrophy cell models using real-time quantitative PCR, enzyme-linked immunosorbent assay or immunoblotting experiments. The correlation of the three molecular expressions was preliminarily verified in AC16 cells. Finally, silencing BPTF, lncRNA TNRC6CAS1 or overexpressing miR-30c-1-3p could inhibit the expression of cardiac hypertrophy marker proteins. The above results indicated that the ceRNA network composed of lncRNA TNRC6CAS1, miR-30c-1-3p, and BPTF may participate in regulating the generation of cardiac hypertrophy in HCM. As potential pathogenic molecules of HCM, the three molecules are expected to become new diagnostic and treatment targets.
  • Research Papers
    WEI Xi-Yang, WANG Zheng-Zhou, JI Kai-Yuan
    Chinese Journal of Biochemistry and Molecular Biol. 2022, 38(12): 1714-1725. https://doi.org/10.13865/j.cnki.cjbmb.2022.10.1233
    Melanoma is a malignant tumor that occurs frequently in the skin. Due to its strong invasiveness and poor prognosis, it has always been the researchers' focus. Circular RNAs (circRNAs) are novel endogenous non-coding RNAs, which are widely involved in animal growth and development, cell differentiation, signal transduction and other physiological processes. However, the molecular mechanism of circRNAs in melanoma cells has not been fully analyzed. In this study, mice (C57BL/6J) normal melanocytes and B16 melanoma cells were used as research objects, and the expression characteristics of circRNAs between the two kinds of cells were analyzed by second-generation sequencing technology. Sequencing results showed that there were 851 circRNAs and 195 differentially expressed circRNAs (DECs) in mouse normal and melanoma cells. GO and KEGG database annotation found that the source genes of DECs were mainly involved in the cell cycle, the tight junction, the Rap1 signaling pathway, the TGF-beta signaling pathway and other signaling pathways related to cell proliferation and migration. CircE2F5 (circ-3:14578602|14606309) was significantly overexpressed in melanoma cells compared with normal melanocyte, which could increase E2F5 expression, promote the melanoma cell proliferation. TargetScan 7.0 and RegRNA 2.0 predicted that there were 73 DECs in mouse normal melanocytes and melanoma cells that had miRNA binding sites, and multiple potentially targeted miRNAs were involved in the proliferation and migration of melanocytes. In sum, multiple DECs related to melanoma cell proliferation and migration were obtained by functional annotation of DEC source genes and potential targeted miRNAs of DECs, and we expected it to provide new insights for melanoma research.
  • Research Papers
    WANG Xiao-Yong, SHANG Ge-Chu, GAO Yan
    Chinese Journal of Biochemistry and Molecular Biol. 2022, 38(11): 1529-1537. https://doi.org/10.13865/j.cnki.cjbmb.2022.09.1214
    Statins can bring some benefits to the treatment of diabetic cardiomyopathy (DCM), but the specific molecular pathway of their action is still unclear. Recent studies have shown that abnormal expression of long noncoding RNA (lncRNA) is closely related to the pathological development of DCM. To compare the degree of myocardial injury between diabetic rats treated with rosuvastatin and rats treated with conventional therapy, the therapeutic pathway and potential target of rosuvastatin on DCM was investigated. Total RNA of DCM rats was extracted and 1ncRNA microarray was prepared to screen out differentially expressed 1ncRNA and bioinformatics analysis was carried out. The results showed that 770 target genes were up-regulated and 884 were down-regulated in the treatment group compared with the model group, which were mainly related to improvement of metabolic disorder, regulation of the ratio of myocardial cells to collagen fibers, reduction of myocardial injury and exercise burden, prevention of autonomic nervous system and microcirculation diseases and change of eating habits. The signaling pathways involved are mainly concentrated in sensory pathways, signal transduction, lipid metabolism and so on. It is suggested that rosuvastatin may play a role in the treatment of DCM by regulating the participation of 1ncRNA in glucose and lipid energy metabolism and ion balance, inhibiting the process of myocardial fibrosis and improving the effect of high glucose toxicity on autonomic nervous function.
  • Research Papers
    AN Xiao-Qiong, XIE Peng, ZHU Xiao-Xi, LONG Ting-Ting, YU Wen-Feng
    Chinese Journal of Biochemistry and Molecular Biol. 2022, 38(10): 1403-1417. https://doi.org/10.13865/j.cnki.cjbmb.2022.08.1207
    In this study, we performed high-throughput sequencing technology methylated RNA immunoprecipitation sequencing (MeRIP-seq), transcriptome sequencing (RNA-seq) and bioinformatics to analyze the differentially m6A-methylated and differentially expressed profile of circular RNA (circRNA) in middle cerebral artery occlusion / reperfusion (MCAO/R) model, which provided some scientific evidences for revealing the relationship between RNA epigenetic modification and cerebral ischemia reperfusion injury. The neurological deficit scores of mice were evaluated by the Longa score standard. TTC staining was used to detect cerebral infarction volumes, and dot blot was used for the quantification of m6A abundance. The results showed that the MCAO/R group showed severe neurological loss, and their Longa scores (2.75 ± 0.25) were significantly increased compared with that in the sham group. The percentage of cerebral infarct volumes in the MCAO/R group (27.63% ± 4.24%) was higher than that in the sham group and cerebral ischemia-reperfusion elevated global m6A levels. Compared with the sham group, 1787 circRNAs in the MCAO/R group were significantly changed, of which 852 circRNAs were increased and 935 circRNAs were decreased significantly (P<0.05). Gene ontology (GO) function analysis showed that differentially target genes were mainly involved in translation, protein N-linked glycosylation, response to hormone, interleukin-6-mediated signaling pathway, Golgi cisterna membrane, and integral component of endoplasmic reticulum membrane, etc. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis found that differential circRNAs were mainly related to N-Glycan biosynthesis, Wnt signaling pathway, gastric acid secretion, glutamatergic synapse, phospholipase D signaling pathway, taste transduction, african trypanosomiasis, thyroid hormone synthesis, insulin secretion and so on. Compared with the sham group, 22 m6A peaks in the MCAO/R group were significantly changed, of which 14 m6A peaks were significantly upregulated and 8 m6A peaks were decreased significantly(P<0.05). GO and KEGG pathway analyses of the predicted target genes of the differentially m6A-methylated circRNAs were performed. GO analysis showed that differentially m6A-methylated target genes were mainly related to 5-methylcytosine catabolic process, chemical homeostasis within a tissue, G-protein coupled glutamate receptor binding and so on. KEGG pathway enrichment analysis indicated that the differentially m6A-methylated circRNAs target genes were mainly related to Fanconi anemia pathway. By comprehensively analyzing MeRIP-seq and RNA-seq data, there were seven differentially m6A-methylated and expressed circRNAs, and RT-qPCR was used to detect their expression. The target genes of the differentially m6A-methylated and differentially expressed circRNAs were predicted. In this study, sequencing analysis of the differentially m6A-methylated and differentially expressed circRNAs from sham and MCAO/R groups, and the results suggest that cerebral ischemia-reperfusion can cause changes of the circRNA expression profile and m6A modification profile in mice, and target genes of differentially expressed circRNA and differentially methylated circRNA are involved in multiple functions and pathways. This study lays the foundation for revealing the molecular mechanism of cerebral ischemia-reperfusion injury from epigenetic level and provides potential target sites for subsequent research on cerebral ischemia-reperfusion injury.
  • Research Papers
    YANG Wan-Yun, AJAB Khan, JIA Qiong, HU Shi-Xiong, JIAO Ding-Xing, YU Lei-Tao, FAN Rui-Wen
    Chinese Journal of Biochemistry and Molecular Biol. 2022, 38(6): 799-808. https://doi.org/10.13865/j.cnki.cjbmb.2022.04.1560
    Long non-coding RNA (lncRNA) is a type of non-coding RNA with the more than 200 nucleotides. Several lncRNAs have been identified as the potential targets for cancer therapy. LncRNA00067110 is one of the differentially expressed genes in the transcriptome profiles of melanoma B16-F10 cells compared to normal mice melanocytes. To investigate whether lncRNA00067110 regulates the proliferation, apoptosis and melanogenesis of B16-F10 cells, the calcium-binding tyrosine phosphorylation regulated protein (Cabyr) target gene was predicted by LncTar and verified by dual luciferase activities. The regulating function of lncRNA00067110 was investigated by the analysis of transcriptome profiles and to detect the proliferation, apoptosis and melanin production of B16-F10 cells transfected by the overexpression plasmids of lncRNA00067110. The results showed that the relationship of lncRNA00067110 targeting Cabyr, the mRNA and protein levels of proliferation (MEK/ERK/ MNK/ CREB) and melanogenesis-related genes (TYR family and CREB) were significantly down-regulated, while the mRNA and protein levels of apoptosis-related genes (AKT and Bcl-2) were up-regulated in B16-F10 cells with lncRNA00067110 overexpression. The transcriptome profile of B16-F10 cells with lncRNA00067110 overexpression showed that 17 genes were differentially expressed, among which Cabyr was up-regulated. Furthermore, the effect of lncRNA00067110 on the phenotypes of cell proliferation and apoptosis were verified. The results suggested that lncRNA00067110 might be a novel target for the treatment of melanoma by targeting Cabyr, which regulate the expression of related genes to inhibit the proliferation and melanogenesis, as well as to induce the apoptosis of B16-F10 cells.
  • Reviews
    HUANG Jiong-Jie, CHEN Le-Yi, LIAO Qi
    Chinese Journal of Biochemistry and Molecular Biol. 2022, 38(4): 438-451. https://doi.org/10.13865/j.cnki.cjbmb.2021.09.1121
    With the development of sequencing technology and in-depth research on tRNA-derived small molecules (tsRNAs), more and more tRNAs and their functions have been identified in various species. tsRNAs can be divided into tRNA-derived fragment (tRF) and tRNA-derived stress-induced RNA (tiRNA) according to different cleavage sites. And we will focus on tRF, which is a kind of non-coding RNA with regulatory function. To deepen the research of tRF, a large number of tRF identification methods based on sequencing data and corresponding databases are being constructed in recent years. The former mainly includes the method of Telonis et al. and tDRmapper, while the latter mainly includes tRFdb, tRF2Cancer and MINTbase. At the same time, both provide a more effective tool for the in-depth research of tRF. The regulation mechanisms of tRF are also being illustrated in many studies. tRF mainly regulates the expression of RNA, DNA and proteins in a miRNA-like manner. With further investigations, researchers have found that tRF also plays a specific regulatory role in various biological processes of human diseases, suggesting its role as a potential biomarker. Herein we mainly summarize the identification methods, databases, regulation mechanisms of tRF and its role in human diseases.
  • Research Papers
    ZHANG Na, LI Meng, LI Jiao, MENG Shan, CAI Chun-Bo, YANG Yang, GAO Peng-Fei, GUO Xiao-Hong, CAO Guo-Qing, LI Bu-Gao
    Chinese Journal of Biochemistry and Molecular Biol. 2022, 38(3): 333-342. https://doi.org/10.13865/j.cnki.cjbmb.2022.02.1554
    Circular RNA (circRNA), as a competitive endogenous RNA (ceRNA), plays an important role in the regulation of cell differentiation. The purpose of this study was to identify and analyze porcine circular RNA insulin-like growth factor 1 receptor (circIGF1R), explore its expression patterns, construct a ceRNA regulatory network related to circIGF1R, and explore the regulation of its ectopic expression on adipogenic differentiation of mouse mesenchymal stem cells (C3H10T1/2) effect. Forward and reverse PCR, Sanger sequencing, RNase R enzyme digestion tests, and qRT-PCR were used to verify that circIGF1R is a circRNA formed by the second exon of insulin-like growth factor 1 receptor (IGF1R). It was expressed in all tissues of pigs, and its expression level increased with age in adipose tissues. miRDB, TargetScan and miRWalk online software were used to predict circIGF1R target genes. RNAhybrid software was used for binding site prediction. DAVID bioinformatics functional analysis software was used to perform GO and KEGG enrichment analysis on candidate target genes. Cytoscape software was used to construct the ceRNA network diagram. Based on the gene expression correlation and predicted target relationship, the GO and KEGG enrichment analysis was drawn and the ceRNA network was constructed; the dual luciferase reporter gene test was used, and we found that circIGF1R and FABP4 can bind to ssc (Sus scrofa chromosome) -miR-133a-5p. The circIGF1R overexpression vector was successfully constructed and expressed in C3H10T1/2 cells. It was found that after overexpression of circIGF1R, the expression of key adipogenic regulatory factors CEBPα, CEBPβ, FABP4 and PPARγ increased significantly(P<0.01), and the number of lipid droplets increased significantly. The results of this study show that circIGF1R exists in pig adipose tissues, and may positively regulate the adipogenic differentiation of C3H10T1/2 cells through the ceRNA mechanism, which lays a theoretical foundation for further research on circIGF1R regulating the adipogenic differentiation of pig precursor intramuscular adipocytes.
  • Research Papers
    LONG Pei-Yan, ZHANG Wen-Ping, ZHENG Ju, FU Yan-Lin, GAO Xiao, WANG Zheng-Wei, XIAO Yan
    Chinese Journal of Biochemistry and Molecular Biol. 2022, 38(3): 356-364. https://doi.org/10.13865/j.cnki.cjbmb.2022.02.1589
    In this study, high-throughput sequencing technology was used to analyze the differentially expressed microRNA (miRNA) of astrocyte-derived exosomes in control group and oxygen and glucose deprivation/reoxygenation (OGD/R) group. Ultracentrifugation was used to extract exosomes from the supernatant of astrocyte medium in the control group and OGD/R group. Transmission electron microscopy showed that exosomes had a typicalvesicle shape with intact membrane and low electron content density. Nanoparticle tracking technology (NTA) detected astrocyte exosomes with a size of 100.5±31.1 nm, accounting for 96.8%. Western blot detection showed that the exosome contained exosome-specific proteins tumor-susceptibility protein (TSG101), heat shock protein 60 (Hsp60), ALG-2-interacting protein X (ALIX). Compared with the control group, 41 miRNAs in the exosomes of the OGD/R group were significantly changed, of which 20 miRNAs were increased and 21 miRNAs were decreased significantly (P<0.05). Gene ontology function (GO) analysis showed that significantly differentially target genes were mainly involved in protein glycosylation, lipid metabolism, phosphorylation, Golgi apparatus, endoplasmic reticulum, endosome, cytoplasmic vesicles and cell protrusions , etc. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis found that differential miRNAs were mainly related to metabolic pathways and signaling pathways such as butyrate metabolism, β-alanine metabolism, fatty acid degradation, mitophagy and P53 signaling pathway. Sequencing analysis of the exosomal miRNAs derived from control and OGD/R astrocytes and target gene function enrichment analysis can be useful for the mechanism study of astrocyte exosomes in response to oxygen and glucose deprivation reperfusion.
  • Non-coding RNA Special Column
    HUA Min-Min, SHI Hui-Juan, RU Yan-Fei
    Chinese Journal of Biochemistry and Molecular Biol. 2022, 38(2): 127-136. https://doi.org/10.13865/j.cnki.cjbmb.2021.09.1139
    Developmental programming of the embryo is controlled by both genetic information and epigenetic information. During fertilization, this information carried by sperms can be delivered to the zygote, where they can regulate early embryonic development. Mature sperms are highly abundant in epigenetic information, and including small non-coding RNAs (sncRNAs), which play important roles during spermatogenesis, fertilization, and early embryo development. Recent studies revealed that sncRNAs can regulate gene expression, mediate protein translation, transmit the epigenetic information, and so on. Recently, increasing evidences showed that parental environment exposure, such as diet, toxicant, pressure, may cause the inheritance of acquired characteristics, and they can be stored and transmitted to the next generation by epigenetic information in germ cells. Recent advances of transgenerational inheritance revealed that sncRNAs are environmentally responsive epigenetic molecules in sperms. This review summarized current knowledge about the sncRNAs information in sperms, including transfer RNA-derived small RNAs (tsRNAs), rsRNAs (risbosome-RNA derived small RNAs), microRNAs (miRNAs) and PIWI-interacting RNAs (piRNAs), that are responsive to environmental factors and are capable of affecting embryonic development and the phenotype of the offspring later in life. Furthermore, this review also delineated potential molecular mechanisms that might regulate sperm sncRNAs.
  • Non-coding RNA Special Column
    GENG Xiu-Chao, LI Qiang, WANG Hong
    Chinese Journal of Biochemistry and Molecular Biol. 2022, 38(2): 137-149. https://doi.org/10.13865/j.cnki.cjbmb.2021.05.1096
    As an important member of the non-coding RNA family, circRNA is a kind of single-stranded RNA with a covalently closed loop structure without a polyadenylated acid tail and 5′-3′ end, showing high stability, abundance and conservation across species characteristics. Recent studies have shown that circRNA plays an important role in many biological processes, including chemotherapy resistance and malignant progression. Exosomes are small extracellular phospholipid bilayer vesicles with a diameter of 30-150 nm that are secreted by living cells. They can be used as carriers to encapsulate and transfer functional molecules. Exosomes are important mediators of communication between tumor cells and stromal cells. They can play a role in the transmission of chemoresistance by transferring circRNA. As chemotherapy resistance is still a huge obstacle to the prognosis of cancer, the research of exosomal circRNA-mediated tumor chemotherapy resistance is at the forefront of academic research, which is a blue ocean with important significance. In this paper, we summarized the latest research progress in the aspects of exosome delivery of circRNA, the mechanism of exosome sorting non-coding RNA cargo, circRNA-mediated tumor chemotherapy resistance, exosome delivery of circRNA-mediated tumor chemotherapy resistance and its potential clinical application, which may provide a reference for the research of tumor chemotherapy resistance.
  • Non-coding RNA Special Column
    LIN Hong, LONG Fang-Yi, WANG Ting
    Chinese Journal of Biochemistry and Molecular Biol. 2022, 38(2): 150-158. https://doi.org/10.13865/j.cnki.cjbmb.2021.05.1033
    Circular RNA (circRNA) is a novel RNA with circular structures. It is conserved in various species, and characterized by high stability, high expression levels and tissue specificity. Meanwhile, it could serve as microRNA (miRNA) sponges, bind to RNA-binding proteins, or regulate transcription and protein translation. With the development of high-throughput sequencing and bioinformatics, circRNA was reported to participate in the pathogenesis of cancer. N6-methyladenosine (m6A) modification is the most common type of RNA modification in eukaryote RNA, which is a dynamic and reversible process regulated by m6A methyltransferase, m6A demethylase and m6A-binding proteins. M6A modification is involved in the regulation of RNA nuclear export, splicing, stability, translation and degradation, thus playing a key role in the occurrence and development of multiple human diseases, such as cancers, cardiovascular diseases. Recently, some researches demonstrated that m6A modification of circRNA was essential in the occurrence and development of malignant tumors, such as cervical cancer, colorectal carcinoma, hepatocellular carcinoma, non-small cell lung cancer, poorly differentiated adenocarcinoma of the stomach. In the current manuscript, we summarized the mechanism of m6A RNA modification, the roles of m6A modification in regulation of circRNA, and the effects of circRNA m6A modification on tumor. The potential clinical application value of m6A-modified circRNA was further discussed, as to provide some new ideas and ways for early diagnosis, clinical treatment and prognosis of tumors.
  • Non-coding RNA Special Column
    YUAN Feng-Ying, SUN Shao-Kang, JIN Zhi-Sheng
    Chinese Journal of Biochemistry and Molecular Biol. 2022, 38(2): 159-164. https://doi.org/10.13865/j.cnki.cjbmb.2021.11.1431
    Diabetes is a clinical syndrome caused by a variety of factors. It often causes multiple system damage, leading to chronic progressive lesions of the eye, kidney, blood vessels, heart, nerves and other organs. At present, its etiology and pathogenesis are not fully clarified, and there is a lack of effective cure. Further exploration of the molecular regulatory mechanisms that drive diabetes and its complications, identifying specific biomarkers and molecular therapeutic targets, is undoubtedly an effective strategy to prevent the onset and development of diabetes and improve the quality of life of patients. Long non-coding RNA (lncRNA) is an important regulator of body normal activity and disease development. Abnormal expression and mutation are one of the main causes of diabetes and many other diseases. Nuclear paraspeckle assembly transcript 1 (NEAT1) is a newly discovered lncRNA molecule in recent years. It has attracted much attention because of its important regulatory role and diverse biological effects in diabetes and its complications. This article summarizes the molecular regulation mechanism and related biological functions of lncRNA NEAT1 in diabetes and its complications in order to provide a new scientific reference for early prevention, diagnosis and molecular targeted therapy of diabetes.
  • Non-coding RNA Special Column
    PAN Jian-Feng, ZHANG Yan-Jun, LI Jin-Quan
    Chinese Journal of Biochemistry and Molecular Biol. 2022, 38(2): 165-174. https://doi.org/10.13865/j.cnki.cjbmb.2021.11.1358
    Long non-coding RNA (lncRNA) are non-coding RNA (ncRNA) greater than 200nt in length, which were initially considered as transcriptional "junk" with no biological function. As research progressed, lncRNA were found to be involved in many biological regulatory processes, such as chromosome silencing, chromatin modification, transcriptional activation and interference. These biological regulatory processes are closely related to the structure and spatial and temporal specific expression of lncRNA. In addition, the corresponding regulatory mechanisms of lncRNA with different structures and locations are different. When lncRNA are located in the nucleus, they mostly regulate gene expression at the transcriptional level and epigenetically, including histone modifications, DNA methylation, chromosome remodeling and other modification processes. In contrast, lncRNA in the cytoplasm mostly play regulatory roles at the post-transcriptional and translational levels, and the mechanisms of action and functions of lncRNA vary among different organelles, all of which illustrate the importance of the location of lncRNA on their functional performance. In addition, exosomes are also rich in lncRNA, and these lncRNA can be delivered to the corresponding sensitive cells through intercellular communication to generate the corresponding regulatory mechanisms. In addition, aberrant expression of lncRNA in different structures is often a key factor in the development and progression of related diseases and cancers. Studying the enrichment of lncRNA in different subcellular structures can help understand the specific roles played by lncRNA in regulating body homeostasis, disease and cancer occurrence and development, as well as growth and development of plants and animals, as well as provide a new theoretical basis for subsequent studies on targeted therapies and improving animal production performance. This paper outlines the latest research progress on the different regulatory mechanisms of lncRNA in chromosomes, membraneless substructures, cytoplasm (endoplasmic reticulum, ribosomes, mitochondria, lysosomes), exosomes and other locations, as well as describes the processes of related diseases and cancers caused by lncRNA abnormalities within the corresponding structures. Finally, an outlook on lncRNA research is given with the aim of providing a corresponding theoretical basis for future studies.
  • Non-coding RNA Special Column
    LI Ke-Xin, TIAN Tong-Tong, ZHOU Bo
    Chinese Journal of Biochemistry and Molecular Biol. 2022, 38(2): 175-181. https://doi.org/10.13865/j.cnki.cjbmb.2021.08.1174
    MicroRNA (miRNA) is a class of endogenous non-coding single-stranded RNA molecules with a length of about 16~29 nucleotides. Widely found in eukaryotes, they play important regulatory roles in plant cell proliferation and differentiation, organ formation, metabolism, resistance to salt, temperature, drought, heavy metal stress, etc. Plant miRNA mainly affects plant growth and development by degrading target genes or inhibiting the expression of target genes at the translation level. At present, the research on the production and regulation of miRNA is relatively clear, and the specific roles and regulatory networks of miRNA in plant secondary metabolism and response to abiotic stress have also been identified, which lays the foundation for a full understanding of the molecular regulation of miRNA. In order to better understand the expression and regulation characteristics of miRNA and to interpret the regulatory network of miRNA in plant secondary metabolism and abiotic stress response, we review the molecular mechanisms of plant miRNAs in regulating the biosynthesis of various secondary metabolites (flavonoids, terpenoids, alkaloids) and responding to environmental stress (salt, high temperature, low temperature, drought, heavy metal stress),and summarize the regulatory roles of miRNA in secondary metabolism and abiotic stress, which will provide references for understanding the relationship between environmental stress and plant metabolism, for further studying the regulatory mechanism of miRNA in maintaining plant homeostasis, and for cultivating superior crop varieties.
  • Non-coding RNA Special Column
    JIA Rui, YAN Shu-Guang, HUI Yi
    Chinese Journal of Biochemistry and Molecular Biol. 2022, 38(2): 182-190. https://doi.org/10.13865/j.cnki.cjbmb.2021.04.1674
    Macrophage polarization is a process of phenotypic regulation based on the surrounding stimulus environment, which is generally polarized into two phenotypes, namely classical activated M1 macrophages and alternative activated M2 macrophages. In short, M1 macrophages are characterized as pro-inflammatory and anti-tumor; M2 macrophages are anti-inflammatory and pro-tumor. Macrophage polarization is considered to be a key regulator of human physiology and pathology processes, and its effectiveness depends on the coordinated expression of key factors, whose expression is finely regulated by microRNAs (miRNAs) at the post-transcriptional level. MicroRNAs are small non-coding RNAs that have the ability to regulate gene expression and cellular network processes. More and more evidence shows that miRNAs play an important role in regulating macrophage polarization. Therefore, in this review, miRNAs regulating macrophage polarization to M1/M2 type and have bidirectional regulation function are listed, and how they regulate macrophage polarization through transcription factors and their potential in the treatment of inflammation and tumor.