miR-21通过调节PTEN及Wnt信号通路介导Huh7细胞对顺铂的耐药性

肝脏 ›› 2018, Vol. 23 ›› Issue (1): 31-36.

miR-21通过调节PTEN及Wnt信号通路介导Huh7细胞对顺铂的耐药性

陆天飞, 钟成鹏, 顾广祥, 郝俊, 张建军, 徐宁

200127 上海交通大学医学院附属仁济医院肝脏外科

收稿日期:2017-11-18 发布日期:2020-04-30
通讯作者: 徐宁,Email:xuning123@hotmail.com
作者简介:共同第一作者:钟成鹏

Regulation of microRNA-21 for cisplatin resistance in Huh7 cells by PTEN and Wnt signaling pathways

LU Tian-fei, ZHONG Cheng-peng, GU Guang-xiang, HAO Jun, ZHANG Jian-jun , XU Ning

Department of Liver Surgery, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200127, China

Received:2017-11-18 Published:2020-04-30
Contact: XU Ning,Email:xuning123@hotmail.com

摘要/Abstract

摘要： 目的探讨microRNAs(miRNAs)介导肝细胞癌(HCC)对顺铂的耐药机制。方法采用miR-21 mimics及miR-21抑制剂来调节Huh7细胞miR-21表达量。人DKK-1与bpv(phen)分别是Wnt 信号通路抑制剂及PTEN抑制剂。相关mRNA及蛋白水平采用RT-PCR及蛋白质免疫检测。结果 miR-21过表达将减弱顺铂(5和10 μg/mL)对Huh7细胞增殖的抑制作用。当Huh7细胞经miR-21 mimics处理后,Wnt4的mRNA及蛋白质表达均显著上升,且miR-21过表达可逆转DKK-1对Wnt4 和细胞增殖的抑制作用。另外,miR-21过表达还上调了PTEN的mRNA及蛋白表达,并可提升经bpv(100和200nmol/L)抑制的Huh7细胞增殖速率。结论 miR-21在Huh7细胞对顺铂的耐药中发挥作用,且通过活化Wnt 信号通路及PTEN调节Huh7细胞增殖。

关键词: 肝细胞肝癌, 顺铂, 化疗耐药性, miR-21, Wnt, PTEN

Abstract: Objective To investigate the mechanism of microRNAs in mediating cisplatin resistance in hepatocellular carcinoma (HCC).Methods MicroRNA-21 (miR-21) level was determined in HCC patients after cisplatin chemotherapy. miR-21 mimics and miR-21 inhibitor were used to upregulate or downregulate miR-21 level in Huh-7 cells. Human dickkopf-1(DKK-1) and bpV (phen) were used as Wnt inhibitor and phosphate and tension homology deleted on chromsome ten (PTEN) inhibitor, respectively. Relative mRNA level and protein expression were detected by real-time polymerase chain reaction (RT-PCR) and western blot, respectively. Results The results showed that overexpression of miR-21 decreased cisplatin (5 and 10 μg/mL)-induced inhibition of cell proliferation in Huh7 cells. Both mRNA level and protein expression of Wnt4 were elevated remarkably when Huh7 cells were treated with miR-21 mimics. Moreover, inhibition of Wnt4 and cell proliferation by DKK-1 was reversed using overexpression of miR-21. In addition, miR-21 mimics increased both mRNA level and protein expression of PTEN, and overexpression of miR-21 increased cell proliferation rate which was inhibited using bpV (phen, 100 and 200 nmol/L) in Huh7 cells. Conclusion miR-21 was involved in regulating resistance to cisplatin in Huh7 cells by activating Wnt and PTEN signaling pathways.

Key words: Hepatocellular carcinoma, Cisplatin, Chemoresistance, miR-21, Wnt, PTEN

陆天飞, 钟成鹏, 顾广祥, 郝俊, 张建军, 徐宁. miR-21通过调节PTEN及Wnt信号通路介导Huh7细胞对顺铂的耐药性[J]. 肝脏, 2018, 23(1): 31-36.

LU Tian-fei, ZHONG Cheng-peng, GU Guang-xiang, HAO Jun, ZHANG Jian-jun , XU Ning. Regulation of microRNA-21 for cisplatin resistance in Huh7 cells by PTEN and Wnt signaling pathways[J]. Chinese Hepatolgy, 2018, 23(1): 31-36.

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