miR-916a调控SOCS6促进HBx-HepG2细胞生长

doi:10.6040/j.issn.1671-7554.0.2016.040

摘要/Abstract

摘要： 目的探讨HBx-HepG2细胞中miR-196a对细胞因子信号抑制物6(SOCS6)表达的调控作用以及对细胞生长的促进作用,并研究其潜在的分子作用机制。方法利用qRT-PCR检测miR-196a以及SOCS6 mRNA的表达水平;Western blotting检测SOCS6蛋白表达水平;CCK-8和集落形成实验检测细胞的生长;采用双荧光素酶报告实验验证miR-196a的靶基因。结果与正常人肝细胞(L02)相比, miR-196a在HBx-HepG2细胞中过量表达(P<0.05)。miR-196a过表达可促进HBx-HepG2细胞生长;miR-196a表达下调可抑制HBx-HepG2细胞生长,与对照组相比差异具有统计学意义(P<0.05)。与正常人肝细胞(L02)相比, SOCS6 mRNA在HBx-HepG2细胞中表达下调(P<0.05);miR-196a可以通过作用于SOCS6的3'UTR区负向调控其表达, 并且SOCS6 的过表达可以抑制HBx-HepG2细胞的生长。结论 miR-196a可以通过靶向负调控SOCS6基因进而促进HBx-HepG2细胞的生长。

关键词: 细胞生长, 细胞因子信号抑制物6, miR-196a, HBx-HepG2细胞

Abstract: Objective To explore the effect of miR-196a on cell growth in HBx-HepG2 cells, and to explore its underlying molecular mechanism. Methods The expressions of miR-196a and SOCS6 mRNA were detected with QRT-PCR, and the protein expression of SOCS6 was determined with Western blotting. The cell growth in HBx-HepG2 cells was evaluated with CCK-8 and colony formation assay. The target gene of miR-196a was validated with dual luciferase reporter experiment. Results The expression of miR-196a was significantly higher in HBx-HepG2 cells than in normal human liver cells(L02)(P<0.05). Over-expression of miR-196a enhanced cell growth in HBx-HepG2 cells(P<0.05), while knockdown of miR-196a suppressed cell growth(P<0.05). SOCS6 was downregulated in HBx-HepG2 cells compared with L02 cells(P<0.05), and miR-196a downregulated the expression of SOCS6 via targeting 3'UTR of SOCS6. Overexpression SOCS6 in HBx-HepG2 cells promoted cell growth. Conclusion miR-196a enhances cell growth via down-regulation of SOCS6 in HBx-HepG2 cells.

Key words: miR-196a, HBx-HepG2 cells, Suppressor of cytokine signaling 6, Cell growth

中图分类号:

R574

余桂芳,陈树娣,陈雪竹,侯开连,梁敏. miR-916a调控SOCS6促进HBx-HepG2细胞生长[J]. 山东大学学报（医学版）, 2016, 54(12): 14-19.

YU Guifang, CHEN Shudi, CHEN Xuezhu, HOU Kailian, LIANG Min. miR-196a promotes cell growth of HBx-HepG2 via targeting SOCS6[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2016, 54(12): 14-19.

参考文献

[1] Khare S, Zhang Q, Ibdah JA. Epigenetics of hepatocellular carcinoma: role of microRNA[J]. World J Gastroenterol, 2013, 19(33): 5439-5445.
[2] Oishi N, Yamashita T, Kaneko S. Molecular biology of liver cancer stem cells[J]. Liver Cancer, 2014, 3(2): 71-84.
[3] Sidhu K, Kapoor NR, Pandey V, et al. The “Macro” world of microRNAs in hepatocellular carcinoma[J]. Front Oncol, 2015, 5: 68. doi: 10.3389/fonc.2015.00068.
[4] Xu Y, Li L, Xiang X, et al. Three common functional polymorphisms in microRNA encoding genes in the susceptibility to hepatocellular carcinoma: a systematic review and meta-analysis[J]. Gene, 2013, 527(2): 584-593.
[5] Hao Y, Wang J, Zhao L. The effect and mechanism of miR196a in HepG2 cell[J]. Biomed Pharmacother, 2015, 72: 1-5. doi: 10.1016/j.biopha.2014.10.032.
[6] Li XD, Li ZG, Song XX, et al. A variant in microRNA-196a2 is associated with susceptibility to hepatocellular carcinoma in Chinese patients with cirrhosis[J]. Pathology, 2010, 42(7): 669-673.
[7] Kokabi N, Xing M, Jr RD, et al. Sociodemographic impact on survival in unresectable hepatocellular carcinoma: a survival epidemiology and end results study[J]. Future Oncol, 2016, 12(2): 183-198.
[8] Buendia MA, Neuveut C. Hepatocellular carcinoma[J]. Cold Spring Harb Perspect Med, 2015, 5(2): a021444. doi: 10.1101/cshperspect.a021444.
[9] Geng M, Xin X, Bi LQ, et al. Molecular mechanism of hepatitis B virus X protein function in hepatocarcinogenesis[J]. World J Gastroenterol, 2015, 21(38): 10732-10738.
[10] 余桂芳, 严跃红, 王瑞鑫, 等. 慢病毒介导HBV X基因稳定表达HepG2细胞系的建立[J]. 世界华人消化杂志, 2012, 20(8): 638-643.
[11] Tsai MM, Wang CS, Tsai CY, et al. MicroRNA-196a/-196b promote cell metastasis via negative regulation of radixin in human gastric cancer[J]. Cancer Lett, 2014, 351(2): 222-231.
[12] Huang F, Tang J, Zhuang X, et al. miR-196a promotes pancreatic cancer progression by targeting nuclear factor kappa-B-inhibitor alpha[J]. PLoS One, 2014, 9(2): e87897. doi: 10.1371/journal.pone.0087897.
[13] Han Q, Zhou C, Liu F, et al. MicroRNA-196a post-transcriptionally upregulates the UBE2C proto-oncogene and promotes cell proliferation in breast cancer[J]. Oncol Rep, 2015, 34(2): 877-883.
[14] Kabir NN, Sun J, Ronnstrand L, et al. SOCS6 is a selective suppressor of receptor tyrosine kinase signaling[J]. Tumour Biol, 2014, 35(11): 10581-10589.
[15] Wu Q, Luo G, Yang Z, et al. miR-17-5p promotes proliferation by targeting SOCS6 in gastric cancer cells[J]. FEBS Lett, 2014, 588(12): 2055-2062.
[16] Wu K, Hu G, He X, et al. MicroRNA-424-5p suppresses the expression of SOCS6 in pancreatic cancer[J]. Pathol Oncol Res, 2013, 19(4): 739-748.
[17] Li ZB, Li ZZ, Li L, et al. miR-21 and miR-183 can simultaneously target SOCS6 and modulate growth and invasion of hepatocellular carcinoma(HCC)cells[J]. Eur Rev Med Pharmacol Sci, 2015, 19(17): 3208-3217.
[18] Qiu X, Zheng J, Guo X, et al. Reduced expression of SOCS2 and SOCS6 in hepatocellular carcinoma correlates with aggressive tumor progression and poor prognosis[J]. Mol Cell Biochem, 2013, 378(1-2): 99-106.
[19] Kazi JU, Sun J, Phung B, et al. Suppressor of cytokine signaling 6(SOCS6)negatively regulates Flt3 signal transduction through direct binding to phosphorylated tyrosines 591 and 919 of Flt3[J]. J Biol Chem, 2012, 287(43): 36509-36517.

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