MiR-126 enhances cisplatin chemosensitivity in hepatocellular carcinoma cells by targeting IRS1

Jian Zuo,Chenjie Huang,Rui Luo,Lanjuan Li,Xiaobin Lou

doi:10.4314/tjpr.v18i1.4

Abstract

Purpose : To investigate the potential role of miR-126 in regulating the proliferation and cisplatin chemosensitivity of human hepatocellular carcinoma (HCC) cells. Methods : The expression of miR-126 was evaluated using clinical HCC specimens. MiR-126-mediated downregulation of Insulin Receptor Substrate 1 (IRS1) was determined by qRT-PCR, western blot and luciferase reporter assay. Cell Counting Kit-8 (CCK8) and colony formation assays were performed to examine the proliferation of HCC cells. Results : Decreased expression of miR126 was found in HCC tumors and was correlated with poor survival in HCC patients. In HCC cells, miR-126 targeted IRS1 for downregulation, through which miR- 126 suppressed the growth of HCC cells and desensitized HCC cells to cisplatin treatment. Conclusion : MiR-126a impairs the proliferation and cisplatin chemoresistance of HCC cells by targeting IRS1. Keywords : miR-126, IRS1, HCC, cisplatin, chemosensitivity

Highlights

Hepatocellular carcinoma (HCC) represents the most common type of liver cancer with approximately 800,000 newly diagnosed HCC cases annually worldwide [1]
Cisplatin, owing to its ability in suppressing tumor growth and inducing tumor cell apoptosis [3,4], is widely used in the treatment of multiple kinds of cancers, including HCC [5].a large proportion of HCC patients can develop cisplatin chemoresistance which leads to the failure of HCC chemotherapy [6]
MicroRNAs are small (19~25 nucleotides in length), non-coding RNAs which bind to the untranslated region (UTR) of target mRNAs, leading to their degradation or plasmid were purchased from Genechem (Shanghai, China)

Summary

INTRODUCTION

Hepatocellular carcinoma (HCC) represents the most common type of liver cancer with approximately 800,000 newly diagnosed HCC cases annually worldwide [1]. Cisplatin, owing to its ability in suppressing tumor growth and inducing tumor cell apoptosis [3,4], is widely used in the treatment of multiple kinds of cancers, including HCC [5].a large proportion of HCC patients can develop cisplatin chemoresistance which leads to the failure of HCC chemotherapy [6]. Total RNA was isolated using TRIzol reagent (Sigma-Aldrich, St. Louis, MO, USA). Tissue samples, including paired tumour and adjacent normal tissues, were collected from 30 HCC patients in The First Affiliated Hospital, College of Medicine, Zhejiang University, between January 2017 and June 2017. The cDNA fragments from IRS1 containing the wild-type (wt) untranslated region (UTR) (IRS13′-UTR-wt) or mutant (mut)-type (IRS1-3′-UTRmut) miR-126 binding sites were cloned into a pGL3 vector (Promega, Sunnyvale, CA, USA).

RESULTS

DISCUSSION

Findings

Conflict of Interest