Small interfering RNA targeting mcl-1 enhances proteasome inhibitor-induced apoptosis in various solid malignant tumors

Xuefeng Huang,Chao He,Wei Zhou,Jingzi Hu,Haimei Tang,Da Wang,Hongbo Zhu

doi:10.1186/1471-2407-11-485

Abstract

BackgroundTargeting the ubiquitin-proteasome pathway is a promising approach for anticancer strategies. Recently, we found Bik accumulation in cancer cell lines after they were treated with bortezomib. However, recent evidence indicates that proteasome inhibitors may also induce the accumulation of anti-apoptotic Bcl-2 family members. The current study was designed to analyze the levels of several anti-apoptotic members of Bcl-2 family in different human cancer cell lines after they were treated with proteasome inhibitors.MethodsDifferent human cancer cell lines were treated with proteasome inhibitors. Western blot were used to investigate the expression of Mcl-1 and activation of mitochondrial apoptotic signaling. Cell viability was investigated using SRB assay, and induction of apoptosis was measured using flow cytometry.ResultsWe found elevated Mcl-1 level in human colon cancer cell lines DLD1, LOVO, SW620, and HCT116; human ovarian cancer cell line SKOV3; and human lung cancer cell line H1299, but not in human breast cancer cell line MCF7 after they were treated with bortezomib. This dramatic Mcl-1 accumulation was also observed when cells were treated with other two proteasome inhibitors, MG132 and calpain inhibitor I (ALLN). Moreover, our results showed Mcl-1 accumulation was caused by stabilization of the protein against degradation. Reducing Mcl-1 accumulation by Mcl-1 siRNA reduced Mcl-1 accumulation and enhanced proteasome inhibitor-induced cell death and apoptosis, as evidenced by the increased cleavage of caspase-9, caspase-3, and poly (ADP-ribose) polymerase.ConclusionsOur results showed that it was not only Bik but also Mcl-1 accumulation during the treatment of proteasome inhibitors, and combining proteasome inhibitors with Mcl-1 siRNA would enhance the ultimate anticancer effect suggesting this combination might be a more effective strategy for cancer therapy.

Highlights

Targeting the ubiquitin-proteasome pathway is a promising approach for anticancer strategies
Rapid accumulation of Mcl-1 induced by proteasome inhibitors in various cancer cells In this study, we evaluated the effect of proteasome inhibitors on Mcl-1 protein expression
Cell lines which were owned by our lab including human colon cancer cell lines DLD1, LOVO, SW620, HCT116, human ovarian cancer cell line SKOV3 and human lung cancer cell line H1299 were treated with different concentrations of bortezomib (0.1-5.0 μM) for 6 h

Summary

Introduction

Targeting the ubiquitin-proteasome pathway is a promising approach for anticancer strategies. Recent evidence indicates that proteasome inhibitors may induce the accumulation of anti-apoptotic Bcl-2 family members. The current study was designed to analyze the levels of several anti-apoptotic members of Bcl-2 family in different human cancer cell lines after they were treated with proteasome inhibitors. The 26S proteasome is a 2, 000-kDa multimeric cylindrical complex comprising a 20S catalytic core and a 19S regulatory subunit [3] This structure is a promising target for cancer therapy because it regulates the crucial process of proteasome-mediated protein degradation, which involves many proteins such as cyclins, caspases, Bcl-2 and the nuclear factor of B (NF-B) [2,4]. The Bcl-2 family proteins, key activators of mitochondrial apoptosis, play a fundamental role in mediating proteasome inhibition-induced toxicity [8].

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