MicroRNA-218下调肝细胞癌中的E2F2基因表达抑制细胞增殖的研究

肝脏 ›› 2016, Vol. 21 ›› Issue (3): 179-182.

MicroRNA-218下调肝细胞癌中的E2F2基因表达抑制细胞增殖的研究

王涛, 马思聪, 戚星星, 汤晓寅, 崔丹, 王智, 池嘉昌, 李萍, 翟博

200127 上海交通大学医学院附属仁济医院肿瘤介入科

收稿日期:2015-10-15 发布日期:2020-07-10
通讯作者: 翟博,Email:zhaiboshi@sina.com
基金资助:
国家自然科学基金(青年项目,81201678);国家自然科学基金(面上项目,81472845)

Study on microRNA-218 inhibiting proliferation of hepatocellular carcinoma cell by down-regulating E2F2 gene expression

WANG Tao, MA Si-cong, QI Xing-xing, TANG Xiao-yin, CUI Dan, WANG Zhi, CHI Jia-chang, LI Ping, ZHAI Bo

Department of Interventional Oncology, Renji Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200127, China

Received:2015-10-15 Published:2020-07-10
Contact: ZHAI Bo, Email:zhaiboshi@sina.com

摘要/Abstract

摘要： 目的探讨MicroRNA-218(miRNA-218)下调肝细胞癌中的E2F2基因表达,从而抑制细胞增殖的机制。方法从细胞库中取得人类肝癌细胞株HepG2、Huh7、MHCC-97H、BEL-7402和正常肝细胞株L02。随后进行细胞培养和转移、反转录多聚酶链反应、免疫印迹分析、细胞增殖和集落形成试验、细胞周期分析和相应的统计学处理。结果 miR-218表达水平在癌细胞中下调。MTT生长曲线表明,当miR-218过度表达时(Huh7中的miR-218 P=0.001;在 MHCC-97中的miR-218 P=0.013)细胞增殖能力明显降低。双荧光素酶实验证实,转染了miRNA-218模拟物的细胞的荧光素酶活性显著降低(P<0.01),荧光酶报告基因含有E2F2野生型的第3′非翻译区。这些结果表明E2F2是 miR-218的直接作用对象。与阴性对照组相比,转染了miR-218 模拟物的Huh7和 MHCC-97细胞株中的E2F2的相对mRNA表达显著下调(P<0.05)。结论 miR-218通过直接作用于E2F2基因的第3′非翻译区调节E2F2的表达,从而抑制HCC细胞增殖。

关键词: MicroRNA-218, 肝细胞癌, E2F2基因, 细胞增殖

Abstract: Objective To investigate the mechanism for microRNA-218 (miR-218) dampening E2F2 gene expression to inhibit proliferation in hepatocellular carcinoma (HCC).Methods HCC cell lines (HepG2, Huh7, MHCC.97H and BEL.7402) and normal hepatocyte cell line L02 were obtained from cell bank, which were subjected for reverse transcription-polymerase chain reaction, immunoblot analysis, cell proliferation and colony formation test, cell cycle analysis and relevant statistics analysis. Results The miR-218 level was down-regulated in cancer cells (P<0.05). The growth curve of MTT indicated that proliferative capacity of cells was significantly reduced in miR-218-overexpressed cells (Huh7: P=0.001; MHCC.97: P=0.013). These results revealed that E2F2 was the direct target of miR-218. In comparison to control group of L02, the relative mRNA of E2F2 in Huh7 and MHCC.97 cell lines transfected with the stimulant of miR-218 was significantly down-regulated (P<0.05). Conclusion miR-218 can directly act on the 3' untranslated region of E2F2 gene to inhibit the development of HCC.

Key words: MicroRNA-218, Hepatocellular carcinoma, E2F2, Cell proliferation

王涛, 马思聪, 戚星星, 汤晓寅, 崔丹, 王智, 池嘉昌, 李萍, 翟博. MicroRNA-218下调肝细胞癌中的E2F2基因表达抑制细胞增殖的研究[J]. 肝脏, 2016, 21(3): 179-182.

WANG Tao, MA Si-cong, QI Xing-xing, TANG Xiao-yin, CUI Dan, WANG Zhi, CHI Jia-chang, LI Ping, ZHAI Bo. Study on microRNA-218 inhibiting proliferation of hepatocellular carcinoma cell by down-regulating E2F2 gene expression[J]. Chinese Hepatolgy, 2016, 21(3): 179-182.

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