Angiogenin (ANG) regulates cell proliferation, migration, differentiation and other biological processes, but the molecular mechanism of ANG function is not fully clear. It has been reported that ANG bound rDNA to enhance rRNA transcription, or might also bind to mRNA coding regions. To screen and identify angiogenin binding sites genome-wide, we employed the technology of chromatin immunoprecipitation coupled with microarray (ChIP-chip) and identified a total of 1 248 binding sites using genomic DNA from HeLa cells. We found 699 potential angiogenin-regulating genes by annotating the genes near the binding sites. Gene annotation and cluster analysis results showed that these genes associated to tumorigenesis and progression linked to TGF-β and Wnt signaling pathway, especially for colorectal and prostate cancers. We verified that angiogenin bound to the promoter regions of the WNT6, CCNE1, APC2, FZD8 and EGFR genes to regulate their expression. These findings provided clues for understanding the mechanisms of angiogenin functions.
Tsuji T, Sun Y, Kishimoto K,et al. Angiogenin is translocated to the nucleus of HeLa cells and is involved in ribosomal RNA transcription and cell proliferation[J].Cancer Res,2005,65(4): 1352- 1360
[2]
Yoshioka N, Wang L, Kishimoto K, et al. A therapeutic target for prostate cancer based on angiogenin-stimulated angiogenesis and cancer cell proliferation[J]. Proc Natl Acad Sci U S A, 2006, 103(39): 14519-14524
[3]
Moroianu J, Riordan JF. Nuclear translocation of angiogenin in proliferating endothelial cells is essential to its angiogenic activity[J]. Proc Natl Acad Sci U S A, 1994, 91(5): 1677-1681
[4]
Xu ZP, Tsuji T, Riordan JF, et al. The nuclear function of angiogenin in endothelial cells is related to rRNA production[J]. Biochem Biophys Res Commun, 2002, 294(2): 287-292
[5]
Li S, Hu GF. Angiogenin-mediated rRNA transcription in cancer and neurodegeneration[J]. Int J Biochem Mol Biol, 2010, 1(1): 26-35
[6]
Xu ZP, Tsuji T, Riordan JF, et al. Identification and characterization of an angiogenin-binding DNA sequence that stimulates luciferase reporter gene expression[J]. Biochemistry,2003, 42(1): 121-128
[7]
Pillai S,Chellappan SP. ChIP on chip and ChIP-Seq assays: genome-wide analysis of transcription factor binding and histone modifications[J]. Methods Mol Biol, 2015, 1288: 447-472
[8]
Negre N, Lavrov S, Hennetin J,et al. Mapping the distribution of chromatin proteins by ChIP on chip[J]. Methods Enzymol, 2006, 410: 316-341
[9]
Furey TS. ChIP-seq and beyond: new and improved methodologies to detect and characterize protein-DNA interactions[J]. Nat Rev Genet, 2012, 13(12): 840-852
[10]
Sheng J, Luo C, Jiang Y, et al. Transcription of angiogenin and ribonuclease 4 is regulated by RNA polymerase III elements and a CCCTC binding factor (CTCF)-dependent intragenic chromatin loop [J]. J Biol Chem ,2014, 289(18): 12520-12534
[11]
Sheng J, Yu W, Gao X, et al. Angiogenin stimulates ribosomal RNA transcription by epigenetic activation of the ribosomal DNA promoter[J]. J Cell Physiol, 2014, 229(4): 521-529
[12]
Luo C, Sheng J, Hu MG, et al. Loss of ARF sensitizes transgenic BRAFV600E mice to UV- induced melanoma via suppression of XPC[J]. Cancer Res, 2013, 73(14): 4337-4348
[13]
Zhu J, Sheng J, Dong H, et al. Phospholipid scramblase 1 functionally interacts with angiogenin and regulates angiogenin-enhanced rRNA transcription[J].Cell Physiol Biochem,2013,32(6): 1695-1706
[14]
Wei S, Gao X, Du J, et al. Angiogenin enhances cell migration by regulating stress fiber assembly and focal adhesion dynamics[J]. PLoS One, 2011, 6(12): e28797
[15]
Polakis P. Wnt signaling in cancer[J]. Cold Spring Harb Perspect Biol , 2012,4(5). pii: a008052. doi: 10.1101/cshperspect.a008052
[16]
Katoh M. WNT signaling in stem cell biology and regenerative medicine[J]. Curr Drug Targets, 2008, 9(7): 565-570
[17]
Senda T, Shimomura A, Iizuka-Kogo A. Adenomatous polyposis coli (Apc) tumor suppressor gene as a multifunctional gene[J]. Anat Sci Int, 2005, 80(3): 121-131
[18]
Sutherland RL, Musgrove EA. Cyclins and breast cancer[J]. J Mammary Gland Biol Neoplasia, 2004, 9(1): 95-104
[19]
Lo HW, Hsu SC, Hung MC. EGFR signaling pathway in breast cancers: from traditional signal transduction to direct nuclear translocalization[J]. Breast Cancer Res Treat, 2006, 95(3): 211-218