Insulin resistance contributes to multidrug resistance in HepG2 cells via activation of the PERK signaling pathway and upregulation of Bcl-2 and P-gp

Liu,Xinyue; Li,Linjing; Li,Jing; Cheng,Yan; Chen,Jing; Shen,Minghui; Zhang,Shangdi; Wei,Hulai

doi:10.3892/or.2016.4632

Insulin resistance contributes to multidrug resistance in HepG2 cells via activation of the PERK signaling pathway and upregulation of Bcl-2 and P-gp

Authors:
- Xinyue Liu
- Linjing Li
- Jing Li
- Yan Cheng
- Jing Chen
- Minghui Shen
- Shangdi Zhang
- Hulai Wei
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Affiliations: Department of Clinical Laboratory Center, The Second Hospital of Lanzhou University, Lanzhou, Gansu 730000, P.R. China, Experimental Center, Northwest University for Nationalities, Lanzhou, Gansu 730000, P.R. China, School of Basic Medical Sciences, Lanzhou University, Lanzhou, Gansu 730000, P.R. China
Published online on: February 24, 2016 https://doi.org/10.3892/or.2016.4632
Pages: 3018-3024

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Abstract

Liver tumorigenesis frequently causes insulin resistance which may be used as an independent risk factor for evaluation of survival and post-surgery relapse of liver cancer patients. In the present study, HepG2/IR, an insulin resistant HepG2 cell line, was established by exposing HepG2 cells to 0.5 µmol/l of insulin for 72 h, and comparison of HepG2/IR with the parental HepG2 cells indicated that the HepG2/IR cells showed significantly enhanced resistance to the most frequently used chemotherapeutics for solid tumors, such as cisplatin, 5-fluorouracil, vincristine and mitomycin. Flow cytometric analysis of cisplatin-treated HepG2/IR cells showed a significantly decreased hypodiploid peak and a significantly downregulated expression level of pro-apoptotic protein caspase-3 compared with the parental HepG2 cells. Our data further showed swollen endoplasmic reticulum (ER) in the cisplatin-treated HepG2/IR cells with significantly increased levels of glucose-regulated protein 78 (GRP78), phosphorylated protein kinase R-like ER kinase (p-PERK) and P-glycoprotein (P-gp). There was also an upregulated expression of anti-apoptotic protein B-cell lymphoma 2 (Bcl-2) whereas no significant change was observed for CCAAT-enhancer-binding protein homologous protein (CHOP), which is known to be induced by ER stress and to mediate apoptosis. Our results demonstrated that insulin resistance in HepG2 cells promoted a protective unfolded protein response and upregulated the expression of ER chaperone protein GRP78, which resulted in the phosphorylation of PERK kinase to activate the PERK-mediated ER stress signal transduction pathway and the upregulation of Bcl-2 and P-gp, leading to the inhibition of the caspase-3-dependent apoptosis pathway and to the survival of liver tumor cells.

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