MicroRNA-539靶向调控E2F转录因子3抑制肝癌的发生与发展

邓学军 龚邵新* 杨胜辉 黄丽荣 周芳

doi:10.16098/j.issn.0529-1356.2018.04.009

PDF(307 KB)

解剖学报 ›› 2018, Vol. 49 ›› Issue (4) : 469-474. DOI: 10.16098/j.issn.0529-1356.2018.04.009

肿瘤生物学

MicroRNA-539靶向调控E2F转录因子3抑制肝癌的发生与发展

邓学军¹ 龚邵新^1* 杨胜辉²黄丽荣³ 周芳³

作者信息 +

MicroRNA-539 inhibiting the occurrence and development of hepatocellular carcinoma by targeting E2F transcription factor 3

DENG Xue-jun¹ GONG Shao-xin ^1* YANG Sheng-hui² HUANG Li-rong³ZHOU Fang³

Author information +

文章历史 +

摘要

目的探讨肝癌组织中microRNA-539（miR-539）表达水平及其抑制肝癌细胞增殖的作用机制。方法收集90例肝癌患者术后肝癌组织及癌旁组织标本，用实时定量聚合酶链反应（Real-time PCR）检测组织中miR-539表达量。在PCL/PRF5、Huh7和QGY-7701肝癌细胞中转染miR-539 模拟物（mimics），用Real-time PCR检测miR-539表达量。用MTT法检测miR-539对肝癌细胞增殖的影响；用流式细胞仪检测miR-539对肝癌细胞凋亡的影响；用免疫印迹法检测miR-539对肝癌细胞中E2F转录因子3（E2F3）蛋白表达的影响。结果肝癌组织中miR-539表达量显著低于癌旁组织（P<0.05）；miR-539表达量与肝癌患者性别、年龄及肝癌组织分化程度无关（P>0.05）；与肿瘤直径和淋巴结转移相关（P<0.05）。转染miR-539 mimics组肝癌细胞中miR-539表达量显著高于空白组和miR-539 NC组（P<0.05）。在QGY-7701细胞中，过表达miR-539能显著抑制细胞增殖（P<0.05），降低E2F3蛋白的表达（P<0.05），对细胞凋亡无明显作用（P>0.05）。结论 miR-539是肝癌发生中的抑癌因子，可能是通过靶向下调E2F3蛋白水平达到抑制癌细胞的增殖。

Abstract

Objective To investigate the expression of microRNA-539 (miR-539) in hepatocellular carcinoma (HCC) and the possible mechanism of miR-539 inhibition cancer cells. Methods Ninety cases of liver cancer patients after liver cancer tissue and adjacent tissue specimens were collected, and the expression of miR-539 in clinical tissues was detected by Real-time PCR. MiR-539 mimics was transfected into QGY-7701, Huh7 and PCL/PRF5 hepatoma cells, and the expression of miR-539 was detected by Real-time PCR. The effect of miR-539 on the proliferation of HCC cells was detected by MTT assay. The effect of miR-539 on the apoptosis of hepatocellular carcinoma cells was detected by flow cytometry. The effect of miR-539 on the expression of E2F transcription factor 3(E2F3) protein in hepatocarcinoma cells was detected by Western blotting. Results The expression of miR-539 in hepatocellular carcinoma was significantly lower than that in adjacent tissues. The expression of miR-539 was not correlated with sex, age and histological differentiation of hepatocellular carcinoma (P>0.05). The expression of miR-539 was correlated with tumor diameter and lymph node metastasis (P<0.05). The expression of miR-539 in miR-539 mimics group was significantly higher than that in blank group and miR-539 NC group (P<0.05). In QGY-7701 cells, overexpression of miR-539 significantly inhibited cell proliferation (P<0.05), decreased E2F3 protein expression (P<0.05), but was no significant effect on apoptosis (P>0.05). Conclusion miR-539 is a tumor suppressor factor in the development of hepatocellular carcinoma, which may inhibit the proliferation of cancer cells through the down-regulation of E2F3 protein level.

导出引用

邓学军龚邵新杨胜辉黄丽荣周芳. MicroRNA-539靶向调控E2F转录因子3抑制肝癌的发生与发展[J]. 解剖学报. 2018, 49(4): 469-474 https://doi.org/10.16098/j.issn.0529-1356.2018.04.009

DENG Xue-jun GONG Shao-xin YANG Sheng-hui HUANG Li-rong ZHOU Fang. MicroRNA-539 inhibiting the occurrence and development of hepatocellular carcinoma by targeting E2F transcription factor 3[J]. Acta Anatomica Sinica. 2018, 49(4): 469-474 https://doi.org/10.16098/j.issn.0529-1356.2018.04.009

参考文献

［1］ Wei KR, Yu X, Zheng RS, et al. Incidence and mortality of liver cancer in China, 2010［J］. Chin J Cancer, 2014, 33(8):388-394.

［2］ Zhang JL, Zhai B, Fang TSh, et al. Progress in the treatment of advanced liver cancer and recurrent liver cancer ［J］. Progress in Modern Biomedicine, 2016, 16 (2): 358-361. (in Chinese)

张金梁, 翟博, 方泰石, 等. 晚期肝癌和复发性肝癌治疗的研究进展［J］. 现代生物医学进展, 2016, 16(2):358-361.

［3］ Altekruse SF, Henley SJ, Cucinelli JE, et al. Changing hepatocellular carcinoma incidence and liver cancer mortality rates in the United States［J］. Am J Gastroenterol, 2014, 109(4):542-553.

［4］ Vitale A, Burra P, Frigo AC, et al. Survival benefit of liver resection for patients with hepatocellular carcinoma across different Barcelona Clinic Liver Cancer stages: a multicentre study［J］. J Hepatol, 2015, 62(3):617-624.

［5］ Fan BL, Ji XH, Zhu WL. Effects of MiR-100 on mitotic arrest and Pololike kinase 1 protein expression in hepatocellular carcinoma cells［J］. Acta Acatomica Sinica, 2014,45(1):70-73. (in Chinese)

范秉琳, 嵇晓辉, 朱武凌. MiR-100对肝癌细胞有丝分裂阻滞及Polo样激酶1蛋白表达的作用［J］. 解剖学报, 2014, 45(1):70-73.

［6］ Pipan Ⅴ, Zorc M, Kunej T. MicroRNA polymorphisms in cancer: a literature analysis［J］. Cancers, 2015, 7(3):1806-1814.

［7］ Reddy KB. MicroRNA (miRNA) in cancer［J］. Cancer Cell Int, 2015, 15(1):1-6.

［8］ Utsunomiya T, Ishikawa D, Asanoma M, et al. Specific miRNA expression profiles of nontumor liver tissue predict a risk for recurrence of hepatocellular carcinoma［J］. Hepatol Res, 2014, 44(6):631-638.

［9］ Gu L, Sun W. MiR-539 inhibits thyroid cancer cell migration and invasion by directly targeting CARMA1［J］. Biochem Biophys Res Commun, 2015, 464(4):1128-1133.

［10］ Nelson HC, Kazuaki C. MicroRNAs as Biomarkers for liver disease and hepatocellular carcinoma［J］. Int J Mol Sci, 2016, 17(3):280.

［11］ Shibata T, Aburatani H. Exploration of liver cancer genomes［J］. Nat Rev Gastroenterol Hepatol, 2014, 11(6):340-349.

［12］ Tai CJ, Huang MT, Wu CH, et al. Contrast-enhanced ultrasound and computed tomography assessment of hepatocellular carcinoma after transcatheter arterial chemo-embolization: a systematic review［J］. J Gastrointestin Liver Dis, 2016, 25(4):499-507.

［13］ Zhang J, Zhang JP. Research progress on hepatocellular carcinoma stem cells［J］. Journal of Hainan Medical University, 2014, 20 (6): 870-872. (in Chinese)

张津, 张俊平. 肝癌干细胞的研究进展［J］. 海南医学院学报, 2014, 20(6):870-872.

［14］ He S, Zhang DC, Wei C. MicroRNAs as biomarkers for hepatocellular carcinoma diagnosis and prognosis［J］. Clin Res Hepatol Gastroenterol, 2015, 39(4):426-434.

［15］ Zeng X, Yin F, Liu X, et al. Upregulation of E2F transcription factor 3 is associated with poor prognosis in hepatocellular carcinoma［J］. Oncol Rep, 2014, 31(3):1139-1146.

［16］ Miles WO, Tsch?p K, Herr A, et al. Pumilio facilitates miRNA regulation of the E2F3 oncogene［J］. Genes Dev, 2012, 26(4):356-368.

［17］ Huang X, Zeng Y, Wang X, et al. FXR blocks the growth of liver cancer cells through inhibiting mTOR-s6K pathway［J］. Biochem Biophys Res Commun, 2016, 474(2):351-356.

［18］ Cao T, Li H, Hu Y, et al. miR-144 suppresses the proliferation and metastasis of hepatocellular carcinoma by targeting E2F3［J］. Tumor Biol, 2014, 35(11):10759-10764.

［19］ Yang Y, Chang S, Zhao Z, et al. MicroRNA-214 suppresses the proliferation of human hepatocellular carcinoma cells by targeting E2F3［J］. Oncol Lett, 2015, 10(6):3779-3784.

［20］ Liu Y, Liang ChH, Li ZZh, et al. The role of microRNA in the occurrence and development of primary hepatocellular carcinoma and its clinical significance［J］. The World Clinical Medicine, 2017,11 (13): 110. (in Chinese)

刘勇, 梁长华, 李增志, 等. MicroRNA在原发性肝细胞肝癌发生与发展中的作用及临床意义［J］. 世界临床医学, 2017, 11(13):110.