Colorectal cancer ranks among the malignancies with high incidence and mortality rates, posing significant challenges to its prevention and treatment. In recent years, accumulating evidence has highlighted the critical involvement of deubiquitinases in the development and progression of colorectal cancer. Deubiquitinases meticulously remove ubiquitin moieties from proteins, thereby regulating protein stability, cellular signaling cascades, and gene expression, which in turn impacts key processes in tumor cells such as proliferation, survival, and metastasis. Deubiquitinases can influence the stability of cell cycle proteins, promoting cell cycle progression and accelerating cellular proliferation. Within the Wnt/β-catenin signaling pathway, deubiquitinases contribute to pathway hyperactivation by enhancing nuclear localization of β-catenin, a pivotal event in colorectal cancer initiation. Deubiquitinases also play a role in modulating the stability of immune checkpoint regulators, affecting the function of immune cells within the tumor microenvironment and facilitating immune evasion. Through regulation of transcription factor ubiquitination status, deubiquitinases impact target gene expression, promoting epithelial-mesenchymal transition, thereby augmenting colorectal cancer's invasive and metastatic potential. Moreover, deubiquitinases mediate chemoresistance in tumor cells by controlling the stability of apoptosis inhibitors, DNA repair enzymes, or drug efflux pumps.Given the critical role of deubiquitinases in colorectal cancer progression, the development of small molecule inhibitors targeting specific deubiquitinases has emerged as an attractive yet challenging field of research. Several inhibitors have demonstrated the capability to inhibit colorectal cancer cell growth and induce apoptosis in vitro and animal models. This review delves into the advancements in understanding the roles of deubiquitinases in colorectal cancer and discusses the application of small molecule inhibitors in colorectal cancer, providing insights for therapeutic strategies against this disease.

Protein methylation is a common post-translational modification in organisms. For a long time, research on protein methylation mainly focused on arginine and lysine, and there were few reports on histidine methylation. However, recent studies have emphasized that histidine methylation is also a widespread and highly conserved modification, occurring at the Nπ and Nτ sites of the histidine imidazole ring, catalyzed by specific protein histidine methyltransferases (PHMTs). Here, we review the history and significant advances in histidine methylation in recent years, particularly highlighting several known histidine methyltransferases. These methyltransferases, through specific molecular mechanisms, are responsible for precise methylation modifications on histidine residues, playing crucial roles in processes such as cell movement, tumor cell proliferation, and protein translation. Additionally, this article discusses the research methods for histidine methylation, especially the application of mass spectrometry, which plays a vital role in advancing histidine methylation research. Although the veil of histidine methylation is gradually being lifted, a complete understanding of this modification and its functional mechanisms still poses challenges. Therefore, this article also presents new insights into the current dilemmas in histidine methylation research and future research priorities, hoping to uncover more secrets of histidine methylation in the future. This could expand the protein methylation modification network and provide new perspectives and strategies for elucidating disease mechanisms and developing new therapeutic approaches.

Depression, a category of mental disorder characterized by core symptoms of anhedonia and depressed mood, seriously influences the physical and psychological health of people worldwide. Clinical and animal studies have been gradually revealing the complicated pathogenic mechanisms involved, and have proposed related hypothesis. The monoamine neurotransmitter deficiency theory has significantly contributed to the development and clinical application of the first-line antidepressants, however, the monoamine targeted drugs generally require more than two weeks of continuous drug treatment, and moreover, are ineffective for approximately one-third of depressed patients. Esketamine is a kind of rapid-acting antidepressant mainly targeting the central glutamatergic system, which is only approved by Food and Drug Administration for treating treatment-resistant depression and major depressive patients with severe suicidal tendency, owing to its potential addictive and psychotomimetic side effects. The search for antidepressants that can rapidly produce effects with minimal side effects remains a key direction for disease treatment, and this endeavor necessitates a deeper understanding of the complex pathogenesis underlying the disease. Recent studies have revealed oxidative stress as a crucial factor in the pathogenesis of depression, and natural antioxidant methods such as exercise and composite dietary can effectively alleviate depressive symptoms, which could serve as promising therapeutic antidepressant approaches. Therefore, we review the close relationship between central glutamatergic system and depression onset, the influence of oxidative stress on glutamate neurotransmission, and the underlying molecular mechanisms involved, to provide novel ideas and drug targets for the prevention and treatment of the disease.

N6-methyladenosine (m⁶A) is a methylation modification on the 6th nitrogen atom of the RNA molecule adenine, and is the most common post-transcriptional modification in messenger RNA (mRNA) and non-coding RNA (ncRNA). m⁶A modification plays a crucial role in all stage of RNA cycle, including RNA stabilization, splicing, nuclear export, folding, translation and degradation. m⁶A modification requires the participation of methyltransferase (writers), demethylase (erasers) and m⁶A readers. The important roles of m⁶A modification involved in virus-host interactions have been largely explored based on the rapid development of high-throughput RNA sequencing technology. Studies have shown that m⁶A modification participates in a variety of viral RNAs, and is associated with viral entrance, replication and progeny virion release. Viruses can also alter host transcriptome by m⁶A modification to impact viral infectivity or host resistance. This review explored the role of m⁶A modification in respiratory virus, retrovirus and herpes virus, etc. infected cells, and the regulatory effects of m⁶A modifications on viral replication and host immune response. It will help to further clarify the mechanisms of virus-host interactions and benefit the development of antiviral drugs.

In natural evolution, cells need to sense various internal and external signals, then process them and execute different functions to adapt environmental changes, the process of which is completed by signal pathways composed of molecules naturally existing in cells. Gene circuits are artificial signal pathways tailored by synthetic biology using engineered or de novo designed biomolecules to realize specific aims like detection or treatment. Gene ciruits consist of 3 components: sensors, prcessors, and actuators. After rational design, gene circuits can sense tumor biomarkers or unbalanced pathological conditions, then release tracer molecules or toxic products to identify or kill tumor cells. These tailored gene circuits exhibited unprecedented specificity in both bacteria and human cells, which has been used to treat cancer in animal experiments. There are 3 approaches when applying gene circuits in cancer treatment: cancer-targeting bacteria based therapies, CAR-T based therapies, and nucleic acid based therapies. Optimization of gene circuits mainly focus on the modularity, orthogonality, adjustability, and composability, in the process of which protein circuits exhibited obvious advantages. They have high reaction rates, excellent composability, and never cause permanent gene changes. The design of gene circuits with multiple sensors and cybernetics-guided design are 2 major research directions of synthetic circuits. Developing sophisticated single transcript protein circuits with multiple sensors and regulators enables new generations of programmable synthetic devices. Here we reviewed the progression of gene circuits in the detection and treatment of cancer, particularly focusing on the application of protease and corresponding cut sites in the design of gene circuits. The flexibility of gene circuit regulation made the design of protein circuits possible, providing ideas for the research of new generation of gene circuits.

RNA m⁶A modification is mainly regulated by m⁶A methyltransferase, m⁶A demethylase and m⁶A binding protein, which can change gene transcription and thus regulating physiological and pathological processes. In recent years, more and more evidences have shown that m⁶A methylation plays an important role in the regulation of tumor microenvironment (TME) . It can affect the occurrence, development and metastasis of various types of cancer. Myeloid-derived suppressor cells (MDSCs), a group of immature myeloid cells, are important immune cells in TME which can be pathologically activated. It mainly inhibits the activity of T cells, so as to promote the immune escape of malignant tumors. Studies have shown that MDSC, a new and promising target for immunotherapy, can reshape the immunosuppressive microenvironment and modulate the efficacy of cancer immunotherapy. The role of m⁶A modification in the activation, differentiation and effector function of some immune cells has been widely concerned. However, the research on how m⁶A modification affects MDSC is still very limited. Therefore, it is particularly important to further explore the relationship between them. Based on the introduction of MDSC and RNA m⁶A modification, this review summarizes the mechanism and research progress of RNA m⁶A modification in regulating MDSC in TME in order to provide new strategies for targeting MDSC from the perspective of epigenetic modification.

In tumor cells, changes in specific amino acids can have a significant impact on the metabolism of cancer cells and alter the response of immune cells in the tumor microenvironment, affecting the occurrence, development, and therapeutic effects of tumors. This article first introduces the role of amino acids in tumors, exploring the metabolism of essential amino acids in tumors from the intake of exogenous amino acids, amino acid modification, and metabolism of branched chain amino acids, and clarify the role of nonessential amino acids in nonessential amino acid deficient tumors. Secondly, the impact of amino acid metabolism reprogramming on the occurrence and development of hepatocellular carcinoma was elucidated. Essential and nonessential amino acids play an indispensable role in hepatocellular carcinoma development. In liver cancer cells, the abnormally high content of essential amino acids promotes the occurrence and development of liver cancer cells, but the increase of branched chain amino acid content has a dual effect. Nonessential amino acids often act as inhibitory factors in liver cancer cells, but arginine can combine with asparagine to form a positive feedback loop and promote the occurrence and development of liver cancer. Finally, the impact of amino acid metabolism reprogramming on the treatment of hepatocellular carcinoma was elucidated. Glutamate, serine, branched chain amino acids, S-adenosylmethionine, glycine, arginine, etc. can serve as targets for liver cancer. Inhibiting the expression of glutarate dehydrogenase can improve the drug sensitivity of liver cancer patients to sorafenib. The reprogramming of amino acid metabolism in the tumor microenvironment of hepatocellular carcinoma promotes the malignant proliferation and immune escape of liver cancer cells. Studying the specific changes in amino acids is beneficial for developing personalized immunotherapy for patients.

In industrial production, the expression level of drug proteins in Chinese hamster ovary cells (CHO) is influenced by many factors: the regulatory elements on transcription and translation, the genomic integration sites, and the expression system. Transcription, as the first step of gene expression, largely affects protein expression, and the promoter plays a crucial role in the initiation of transcription. Most of the promoters were screened through transient transfection or random integration, but the presence of unclear copy number or random integration sites makes it difficult to accurately evaluate the promoter activity. To some extent, site-specific integration can reduce the impact of positional effects on exogenous genes and may potentially increase the expression level of exogenous genes. In the early stage of our research, multiple sites that can stably express exogenous proteins were identified and verified in the CHO cell genome. In this study, one of these sites (2c6) was selected for the evaluation of promoter activity. The CRISPR/Cas9 gene editing technique was used to site-specifically integrate the reporter gene (EGFP) regulated by the simian virus early promoter (SV40), mouse elongation factor-1α (mEF-1α), chicken β-actin (cACTB) promoter, and human phosphoglycerate kinase promoter (hPGK) into the 2c6 site, respectively. The mean fluorescence intensity of the cells was analyzed by flow cytometry, and the mRNA level of EGFP was detected by qPCR to comprehensively evaluate the activity of the promoter. The results showed that the activities of the mEF-1α and mACTB promoters were better than those of SV40 and hPGK. The results of the secondary flow cytometry sorting showed that site-specific integration can more accurately evaluate the activity of the promoter in the CHO cell expression system.

Tamoxifen (TAM) has been widely used for the treatment of ER+ breast cancer. However, the inevitable emergence of resistance to tamoxifen obstructs the successful treatment of this cancer. The tumor suppressor gene N-myc downstream-regulated gene 2 (NDRG2) plays a significant role in the development of ER+ breast cancer. However, it is unclear whether NDRG2 participates in mediating TAM resistance in ER+ breast cancer. Here, we investigate the expression of NDRG2 mRNA and protein in TAM-sensitive and TAM-resistant ER+ breast cancer cells. The results of immunoblotting experiments revealed a negative correlation between NDRG2 expression and TAM resistance ability in ER+ breast cancer cells (P<0.001). CCK-8 cell viability assays and soft agar colony formation assays showed that NDRG2 overexpression in TAM resistant cells significantly reduced the TAM IC₅₀ value and the soft agar colony formation rate (P<0.001). For the mechanism, the ERAD reporter protein assays showed that NDRG2 overexpression upregulated the expression of the ERAD reporter protein CD3ε-YFP and increased the levels of spliced XBP1s mRNA, leading to severe endoplasmic reticulum stress in TAM resistant cells (P<0.001). Immunoblot analysis confirmed that overexpression of NDRG2 significantly increased the level of phosphorylation of the endoplasmic reticulum stress sensor IRE1α and the expression levels of its downstream protein factors, including ERdj4, P58IPK, EDEM and PDIA5 (P<0.001). The in vivo xenograft tumor experiments in mice further verified that NDRG2 overexpression significantly inhibited the growth of resistant tumors, which enhanced the therapeutic effect of TAM (P<0.001). These findings indicate that increasing NDRG2 expression and triggering severe endoplasmic reticulum stress upon TAM treatment can reverse the resistance of ER+ breast cancer cells to TAM and inhibits the growth of ER+ breast cancer tumors. Our results provide valuable new insights and potential targets for improving the clinical management of TAM-resistance and prognosis in ER+ breast cancer.

The aim of this study was to investigate the impact of overexpressing 70-kD heat shock proteins (Hsp70) on glycolysis in C2C12 cells during myogenesis and adipogenesis. Using C2C12 cells as the research material, adenovirus was used to overexpress the Hsp70 gene, and changes in the expression of glycolytic genes were detected using fluorescence quantitative PCR and Western blotting techniques. The study indicated that during C2C12 cell myogenic differentiation, the expression trend of the Hsp70 gene was consistent with that of Gsk3β, Pkm, Prkag3, Pfkm, and Hk-2 genes, suggesting a relationship between Hsp70 and the glycolytic pathway during myogenic differentiation. Overexpression of Hsp70 in the later stages of myogenic differentiation significantly upregulated the expression of Gsk3β, Pkm, Prkag3, and Pfkm genes (P<0.05), with no significant impact on Hk-2 gene expression (P>0.05). During C2C12 cell adipogenic induction, the expression trend of the Hsp70 gene was similar to that of Gsk3β, Pkm, Prkag3, Pfkm, and Hk-2 genes, indicating a relationship between Hsp70 and the glycolytic pathway during adipogenic induction. Following Hsp70 overexpression, in the later stages of adipogenic induction, the number of lipid droplets was significantly higher compared to the control group, with a significant upregulation of Gsk3β, Pkm, Prkag3, and Pfkm gene expression (P<0.05), while Hk-2 gene expression was not significantly affected (P>0.05). In conclusion, Hsp70 in C2C12 cells in myogenic and adipogenic states promoted the breakdown of glycogen into 6-phospho-glucose, thereby enhancing the glycolytic pathway, providing insights into the functional role of the Hsp70 gene in glycolysis in C2C12 cells.

As a unique gene in the genome, FLASH (FADD-like interleukin-1β-converting enzyme associated huge protein)/CASP8AP2 is involved in multiple cellular processes, including apoptosis, histone gene pre-mRNA processing, transcriptional regulation, and cell cycle progression. Clinical studies have shown that FLASH is a valuable prognostic marker for acute lymphoblastic leukemia, and a crucial factor for the survival of various cancer cells. Therefore, in-depth research into the function of FLASH may offer new perspectives for the treatment of related diseases. Our previous research identified FLASH as a binding partner of p53, demonstrating that FLASH enhances the transcriptional activity of p53. Here we further investigate the molecular mechanisms of the interaction between FLASH and p53, revealing that the p53-K386R mutation (SUMOylation residue) attenuated its interaction with FLASH (aa 51-200) and FLASH-SIM (SUMO-interacting motif) (aa 1 534-1 806) significantly. However, SUMO can bind to FLASH-SIM directly, instead of FLASH (aa 51-200). Subsequent research shows that overexpression of FLASH in cells enhances global SUMO1 conjugation and p53-SUMO1 conjugation, therefore providing a plausible explanation for the underlying mechanism of FLASH enhancing the transcriptional activity of p53. Since promyelocytic leukemia protein nuclear body (PML NB) serves as subcellular reactors for SUMO conjugation within the cell, and the PML IV isoform can specifically enhance the SUMO modification of p53, we have investigated the interaction between FLASH and PML IV, and elucidated the structural basis of their interaction: both FLASH-N3A (501-802) and FLASH-C2 (1 807-1 981) bind to PML IV (aa 228-633). Further investigations reveal that they can synergistically enhance global SUMO1 modification as well as SUMO1 modification of p53. The interaction between FLASH and tumor suppressors p53 or PML IV enriches our understanding of its function and reveals the potential mechanism of FLASH in tumor development, therefore offering novel insights into cancer diagnosis and treatment.

All-trans retinoic acid (ATRA) is able to induce promyelocytic differentiation effectively. However, its role in the process of erythroid differentiation remains unclear. To investigate the role of ATRA in the process of erythroid differentiation and its epigenetic regulatory mechanism, we established an induced leukemia cell K562 model in this study. Firstly, hemin was used to induce the differentiation of K562 cells into erythroid cells. The results of flow cytometry showed that ATRA affected the lineage changes of cells during erythroid differentiation and blocked the process of cell differentiation. After ATRA treatment of differentiating cells, the expression level of erythroid differentiation-related genes decreased. Through chromatin conformational capture (3C), formaldehyde-assisted separation of regulatory elements (FAIRE), chromatin immunoprecipitation (ChIP) techniques, the epigenetic mechanism was explored and it was found that after ATRA treatment of cells, the chromatin accessibility within the β-globin family gene locus decreased, and the frequency of interaction between the locus control region (LCR) and its target gene promoter decreased. The decrease in the chromatin accessibility of the gene locus led to a decrease in the enrichment frequency of erythroid-related transcription factors GATA binding protein 1 (GATA1), LIM domain binding 1 (LDB1), LIM domain only 2 (LMO2), and BHLH transcription factor 1 (TAL1) at the promoter regions of the LCR and the gene locus of the globin family. The above results indicate that the ATRA treatment of differentiating cells leads to a decrease in the chromatin accessibility of erythroid differentiation-related genes, and a more closed chromatin structure hinders the binding of LCR-recruiting transcription factors to the promoter regions of genes, thereby further repressing the expression of β-globin family genes. This dynamic process elucidates the epigenetic mechanism of ATRA in regulating erythroid differentiation.

Alzheimer's disease (AD) is a neurodegenerative disease with age-related cognitive decline. Sphingosine-1-phosphate receptor 2 (S1PR2) is involved in a variety of cellular processes and has been shown to play an important role in nervous system development. This study aimed to investigate the effects and possible mechanism of S1PR2 on Aβ_25-35 induced cell model damage of AD. In this study, SH-SY5Y cells were induced by Aβ_25-35 to construct a cell damage model, and the expression of S1PR2 in cells was interfered by targeting sequence. The protein and gene expression levels of S1PR2 were detected by Western blot and RT-PCR. It was found that the expression of S1PR2 was significantly increased at mRNA and protein levels in Aβ_25-35-induced SH-SY5Y cell model (P<0.01), and its expression was significantly decreased after S1PR2 intervention (P<0.001). The cell proliferation activity was detected by CCK8, and apoptosis was detected by flow cytometry. The results showed that the proliferation activity of Aβ_25-35 induced SH-SY5Y cells was significantly increased, and apoptosis was decreased after S1PR2 intervention (P<0.01). The protein levels of APP, Tau, p-Tau, and PSD95 in cells were detected by Western blot to analyze the effect of S1PR2 on the pathology of AD. It was found that after S1PR2 intervention, the expressions of APP, Tau, and p-Tau in the AD cell model were significantly decreased (P<0.001), and the expression of synaptic protein PSD95 was increased (P<0.001), which could significantly improve the pathological damage in Aβ_25-35-induced SH-SY5Y cell model. In addition, ATP production was detected by the kit, and ROS content and mitochondrial membrane potential were detected by flow cytometry to analyze the mitochondrial function. Results found that ATP production and mitochondrial membrane potential was significantly decreased , whereas the ROS content was increased in Aβ_25-35 induced SH-SY5Y cells (P<0.001). Intervention with S1PR2 significantly increased ATP production and mitochondrial membrane potential, but decreased ROS content(P<0.001). Finally, the protein levels of the AKT/mTOR pathway were detected by Western blot. The results showed that S1PR2 significantly inhibited the activation of the AKT/mTOR pathway induced by Aβ_25-35 in SH-SY5Y cells. In conclusion, S1PR2 may be involved in the pathogenesis of Aβ_25-35-induced SH-SY5Y cells by promoting mitochondrial function through the AKT/mTOR pathway.

Interleukin-11 (IL-11) is a multifunctional cytokine that plays a crucial role in various biological processes, including the promotion of hematopoiesis, regulation of the immune system, and facilitation of tissue repair. These functions highlight its significant importance in both medical research and clinical therapy. Consequently, the precise detection and assessment of IL-11′s biological activity are essential. The currently employed cell proliferation inhibition assay is cumbersome, time-consuming, and susceptible to interference from Interleukin-6. Here we study the JAK-STAT signaling pathway, a key mechanism of IL-11 actions. We initially establish a stable monoclonal cell line that expresses a luciferase reporter gene responsive to IL-11. We subsequently optimized critical parameters, including the pre-dilution concentration and gradient of IL-11, cell inoculum size, and IL-11 incubation duration. Comprehensive validation was undertaken to evaluate accuracy, precision, specificity, stability, and concordance with pharmacopeial methods, ultimately leading to the establishment of a novel reporter gene-based approach for detecting the biological activity of IL-11. This innovative method significantly enhances the detection accuracy and sensitivity while reducing the testing time, thereby offering promising prospects for broad applications.

Molecular Biology is a key basic professional course for all the students specializing in Biology, Biotechnology, and Bioengineering. With the promotion of double world-class project and first-class undergraduate construction, the development of English-taught course faces challenges. We started to teach the Molecular Biology course in English at the East China University of Science and Technology since 2019, the construction of Molecular Biology course has been reformed and practiced, including the combination of imagery, vividness and classroom teaching, the combination of advanced, cutting-edge and classical theories, and the comprehensive coverage of the teaching process, which has effectively promoted the construction and practice of Molecular Biology course. The Molecular Biology course taught in English greatly increased the students' professional and scientific research ability, international vision and English academic communication ability, comprehensive ability and satisfaction, and teachers' teaching and research ability. This course provides an effective reference for fostering innovative professional first-class undergraduates and the construction of Molecular Biology course.