Zhiyuan WEI, Gaoxing LUO, Yi TIAN, Bing NI, Xiaohe SHEN, Yi SUN

Contribution of hepatitis B virus X protein-induced aberrant microRNA expression to hepatocellular carcinoma pathogenesis

The hepatitis B virus-encoded X (HBX) protein plays important roles in Hepatocellular carcinoma (HCC). Previous studieshave demonstrated that HBX can induce alterations in the expression of numerous microRNAs (miRNAs) involved in the carcinogenesisof various tumors. However, the global profile of liver miRNA changes induced by HBX has not been characterized. In this study, weconducted a miRNA microarray analysis to investigate the influence of HBX on the expression of total miRNAs in liver in relation toHCC. Comparative analysis of the data from human normal liver cells (L02) and human HCC cells (HepG2), with or without HBX,identified 19 differentially expressed miRNAs, including 5 with known association to HBX. Target gene prediction for the aberrantlyexpressed miRNAs identified a total of 304 potential target genes, involved in sundry pathways. Finally, pathway analysis of the HBXinduced miRNAs pathway showed that 5 of the total miRNAs formed an internetwork, suggesting that HBX might exert its pathologicaleffects on hepatic cells through functional synergy with miRNAs that regulated common pathways in liver cells. Therefore, this workprovides new insights into the mechanisms of HCC as well as potential diagnostic markers or therapeutic targets for use in clinicalmanagement of HCC.

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Ahlqvist K, Saamarthy K, Syed Khaja AS, Bjartell A, Massoumi R (2013). Expression of Id proteins is regulated by the Bcl-3 proto-oncogene in prostate cancer. Oncogene 32:1601-1608.
Bilban M, Buehler LK, Head S, Desoye G, Quaranta V (2002). Normalizing DNA microarray data. Curr Issues Mol Biol 4: 57-64.
Chen HS, Kaneko S, Girones R, Anderson RW, Hornbuckle WE, Tennant BC, Cote PJ, Gerin JL, Purcell RH, Miller RH (1993). The woodchuck hepatitis virus X gene is important for establishment of virus infection in woodchucks. J Virol 67: 1218-1226.
Chiang TS, Yang KC, Chiou LL, Huang GT, Lee HS (2014). Enhancement of CYP3A4 activity in Hep G2 cells by lentiviral transfection of hepatocyte nuclear factor-1 alpha. PloS One 9: e94885. Esteller M (2011). Non-coding RNAs in human disease. Nat Rev Genet 12:861-874.
Fang Y, Sun B, Li Z, Chen Z, Xiang J (2016). MiR-622 inhibited colorectal cancer occurrence and metastasis by suppressing K-Ras. Mol Carcinogene 55: 1369-1377.
Guo XB, Jing CQ, Li LP, Zhang L, Shi YL, Wang JS, Liu JL, Li CS (2011). Down-regulation of miR-622 in gastric cancer promotes cellular invasion and tumor metastasis by targeting ING1 gene. World J Gastroentero 17: 1895-1902.
Kew MC (2010). Epidemiology of chronic hepatitis B virus infection, hepatocellular carcinoma, and hepatitis B virus-induced hepatocellular carcinoma. Pathol Biol 58: 273-277.
Lee AT, Ren J, Wong ET, Ban KH, Lee LA, Lee CG (2005). The hepatitis B virus X protein sensitizes HepG2 cells to UV lightinduced DNA damage. J Biol Chem 280: 33525-33535.
Majano P, Lara-Pezzi E, Lopez-Cabrera M, Apolinario A, MorenoOtero R, Garcia-Monzon C (2001). Hepatitis B virus X protein transactivates inducible nitric oxide synthase gene promoter through the proximal nuclear factor kappaB-binding site: evidence that cytoplasmic location of X protein is essential for gene transactivation. Hepatology 34: 1218-1224.
Merico D, Isserlin R, Stueker O, Emili A, Bader GD (2010). Enrichment map: a network-based method for gene-set enrichment visualization and interpretation. PloS One 5: e13984.
Sells MA, Chen ML, Acs G (1987). Production of hepatitis B virus particles in Hep G2 cells transfected with cloned hepatitis B virus DNA. P Natl Acad Sci USA 84: 1005-1009.
Shirakata Y, Koike K (2003). Hepatitis B virus X protein induces cell death by causing loss of mitochondrial membrane potential. J Biol Chem 278: 22071-22078.
Song WH, Feng XJ, Gong SJ, Chen JM, Wang SM, Xing DJ, Zhu MH, Zhang SH, Xu AM (2015). microRNA-622 acts as a tumor suppressor in hepatocellular carcinoma. Cancer Biol Ther 16: 1754-1763.
Sousa S, Cabanes D, Archambaud C, Colland F, Lemichez E, Popoff M, Boisson-Dupuis S, Gouin E, Lecuit M, Legrain P, Cossart P (2005). ARHGAP10 is necessary for alpha-catenin recruitment at adherens junctions and for Listeria invasion. Nat Cell Biol 7: 954-960.
Srisuttee R, Koh SS, Malilas W, Moon J, Cho IR, Jhun BH, Horio Y, Chung YH (2012). SIRT1 sensitizes hepatocellular carcinoma cells expressing hepatitis B virus X protein to oxidative stressinduced apoptosis. Biochem Bioph Res Co 429: 45-50.
Sun Q, Wang R, Wang Y, Luo J, Wang P, Cheng B (2014). Notch1 is a potential therapeutic target for the treatment of human hepatitis B virus X protein-associated hepatocellular carcinoma. Oncol Rep 31: 933-939.
Tian Y, Yang W, Song J, Wu Y, Ni B (2013). Hepatitis B virus X proteininduced aberrant epigenetic modifications contributing to human hepatocellular carcinoma pathogenesis. Mol Cell Biol 33: 2810-2816.
Trang P, Weidhaas JB, Slack FJ (2008). MicroRNAs as potential cancer therapeutics. Oncogene 27 Suppl 2: S52-57.
Vivanco I, Sawyers CL (2002). The phosphatidylinositol 3-Kinase AKT pathway in human cancer. Nat Rev Cancer 2: 489-501.
Wei X, Tan C, Tang C, Ren G, Xiang T, Qiu Z, Liu R, Wu Z (2013). Epigenetic repression of miR-132 expression by the hepatitis B virus x protein in hepatitis B virus-related hepatocellular carcinoma. Cell Signal 25: 1037-1043.
Wei Z, Gao W, Wu Y, Ni B, Tian Y (2015). Mutual interaction between BCL6 and microRNAs in T cell differentiation. RNA Biol 12: 21-25.
Wu CS, Yen CJ, Chou RH, Chen JN, Huang WC, Wu CY, Yu YL (2014). Downregulation of microRNA-15b by hepatitis B virus X enhances hepatocellular carcinoma proliferation via fucosyltransferase 2-induced Globo H expression. Int J cancer 134: 1638-1647.
Xu JD, Cao XX, Long ZW, Liu XP, Furuya T, Xu JW, Liu XL, De Xu Z, Sasaki K, Li QQ (2011). BCL2L10 protein regulates apoptosis/ proliferation through differential pathways in gastric cancer cells. J Pathol 223: 400-409.
Xu X, Fan Z, Kang L, Han J, Jiang C, Zheng X, Zhu Z, Jiao H, Lin J, Jiang K et al. (2013). Hepatitis B virus X protein represses miRNA-148a to enhance tumorigenesis. J Clin Invest 123: 630- 645.
Xu Y, Huang J, Ma L, Shan J, Shen J, Yang Z, Liu L, Luo Y, Yao C, Qian C (2016). MicroRNA-122 confers sorafenib resistance to hepatocellular carcinoma cells by targeting IGF-1R to regulate RAS/RAF/ERK signaling pathways. Cancer Lett 371: 171-181.
Zauri M, Berridge G, Thezenas ML, Pugh KM, Goldin R, Kessler BM, Kriaucionis S (2015). CDA directs metabolism of epigenetic nucleosides revealing a therapeutic window in cancer. Nature 524: 114-118.
Zhang T, Zhang J, Cui M, Liu F, You X, Du Y, Gao Y, Zhang S, Lu Z, Ye L, Zhang X (2013). Hepatitis B virus X protein inhibits tumor suppressor miR-205 through inducing hypermethylation of miR-205 promoter to enhance carcinogenesis. Neoplasia 15: 1282-1291.
Zhou B, Chen H, Wei D, Kuang Y, Zhao X, Li G, Xie J, Chen P (2014). A novel miR-219-SMC4-JAK2/Stat3 regulatory pathway in human hepatocellular carcinoma. J Exp Clin Canc Res 30: 33-55.
Zhou JN, Zeng Q, Wang HY, Zhang B, Li ST, Nan X, Cao N, Fu CJ, Yan XL, Jia YL et al. (2015). MicroRNA-125b attenuates epithelial-mesenchymal transitions and targets stem-like liver cancer cells through small mothers against decapentaplegic 2 and 4. Hepatology 62: 801-815.