解剖学报

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 • 上一篇    下一篇

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   

  1. 1.南华大学附属第一医院病理科，湖南 衡阳421001; 2.湖南中医药大学医学院免疫学教研室，长沙410208; 3.湖南环境生物职业技术学院妇产科教研室，湖南 衡阳 421005
  • 收稿日期:2017-11-29 修回日期:2017-12-14 出版日期:2018-08-06 发布日期:2018-08-06
  • 通讯作者: 龚邵新 E-mail:dengxuejun368@163.com

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-rong3 ZHOU Fang3   

  1. 1.Department of Pathology, the First Hospital Affiliated to Nanhua University, Hu’nan Hengyang421001, China; 2.Department of Immunology, Hu’nan University of Chinese Medicine Medical School, Changsha410208, China; 3.Department of Obstetrics and Gynecology, Hu’nan Vocational College of Environmental Biology, Hu’nan Hengyang421005, China
  • Received:2017-11-29 Revised:2017-12-14 Online:2018-08-06 Published:2018-08-06
  • Contact: GONG Shao-xin E-mail:dengxuejun368@163.com

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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。

关键词: 肝癌, 微小RNA-539, 转录因子, 增殖, 实时定量聚合酶链反应, 免疫印迹法, 人

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.

Key words: Hepatocellular carcinoma, microRNA-539, Transcription factor, Proliferation, Real-time PCR, Western blotting, Human