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Advanced Pharmaceutical Bulletin
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Article History
Submitted: 27 Nov 2013
Revised: 01 Jan 2014
First published online: 07 Feb 2014

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Adv Pharm Bull. 2014;4(3):243-248 doi: 10.5681/apb.2014.035
PMID:24754007        PMCID:PMC3992959

Therapeutic Effects of Myeloid Cell Leukemia-1 siRNA on Human Acute Myeloid Leukemia Cells

Original Research

Hadi Karami 1,2, Behzad Baradaran 1 * , Ali Esfahani 3, Masoud Sakhinia 4, Ebrahim Sakhinia 5,6 *

1 Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
2 Department of Biochemistry, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.
3 Hematology and Oncology Research Center, Shahid Ghazi Hospital, Tabriz University of Medical Sciences, Tabriz, Iran.
4 Faculty of Medicine, University of Liverpool, Liverpool, United Kingdom.
5 Department of Genetics, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.
6 Tuberculosis and Lung Disease Reseach Center, Tabriz University of Medical Sciences, Tabriz, Iran.



Abstract
Purpose: Up-regulation of Mcl-1, a known anti-apoptotic protein, is associated with the survival and progression of various malignancies including leukemia. The aim of this study was to explore the effect of Mcl-1 small interference RNA (siRNA) on the proliferation and apoptosis of HL-60 acute myeloid leukemia (AML) cells. Methods: siRNA transfection was performed using Lipofectamine™2000 reagent. Relative mRNA and protein expressions were quantified by quantitative real-time PCR and Western blotting, respectively. Trypan blue assay was performed to assess tumor cell proliferation after siRNA transfection. The cytotoxic effect of Mcl-1 siRNA on leukemic cells was measured using MTT assay. Apoptosis was detected using ELISA cell death assay. Results: Mcl-1 siRNA clearly lowered both Mcl-1 mRNA and protein levels in a time-dependent manner, leading to marked inhibition of cell survival and proliferation. Furthermore, Mcl-1 down-regulation significantly enhanced the extent of HL-60 apoptotic cells. Conclusion: Our results suggest that the down-regulation of Mcl-1 by siRNA can effectively trigger apoptosis and inhibit the proliferation of leukemic cells. Therefore, Mcl-1 siRNA may be a potent adjuvant in AML therapy.





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