Affiliations 

  • 1 Department of Human Anatomy, Faculty of Medicine and Health sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia
  • 2 Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran
  • 3 Institute of Biosciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia
  • 4 Department of Surgery, Imam Reza Hospital, Tabriz University of Medical Sciences, Tabriz, Iran
  • 5 Neurosciences Research Center, Tabriz University of Medical Sciences, Tabriz, Iran
J Biol Regul Homeost Agents, 2016 Jan-Mar;30(1):55-65.
PMID: 27049076

Abstract

A key issue in the treatment of acute myeloid leukemia (AML) is the development of drug resistance to chemotherapeutic agents. Overexpression of myeloid cell leukemia-1 (Mcl-1), an anti-apoptotic protein, is associated with tumor progression and drug resistance in leukemia and several cancers. The purpose of this study was to investigate the effect of specific Mcl-1 small interference RNA (siRNA) on the proliferation and chemosensitivity of U-937 AML cell to etoposide. The siRNA transfection was conducted using Lipofectamine™ 2000. Quantitative real-time RT-PCR (qRT-PCR) and Western blot analysis were employed to measure the expression levels of mRNA and protein, respectively. To evaluate tumor cell growth after siRNA transfection, Trypan blue exclusion assay was conducted. The cytotoxic effects of siRNA and etoposide were determined using MTT assay on their own and in combination. DNA-histone ELISA and annexin-V/FITC assays were performed to study the apoptosis. Mcl-1 siRNA transfection significantly blocked the expression of Mcl-1 mRNA and protein in a time-dependent manner, leading to a strong growth inhibition and enhanced apoptosis (P less than 0.05). Furthermore, pretreatment with Mcl-1 siRNA, synergistically enhanced the cytotoxic and apoptotic effects of etoposide (P less than 0.05). Our results demonstrated that Mcl-1 plays a fundamental role in the survival and resistance of U-937 cells to etoposide. Therefore, Mcl-1 can be considered an attractive target in gene therapy of AML patients and siRNA-mediated silencing of this gene may be a novel strategy in AML treatment.

* Title and MeSH Headings from MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.