miR-1266-5p and miR-185-5p Promote Cell Apoptosis in Human Prostate Cancer Cell Lines

Document Type: Research Articles

Authors

1 Department of Biochemistry, Pasteur Institute of Iran, Tehran, Iran.

2 Department of Laboratory Medicine, Department of Experimental Cancer Medicine, Karolinska Institutet Huddinge, 141 86 Stockholm, Sweden.

3 Molecular Genetics and Microbiology, University of Toronto, Toronto, Ontario, Canada.

4 Department of Molecular Medicine, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran.

Abstract

Objective: Small non-coding RNA molecules are dysregulated in prostate cancer (PCa). In our previous study,
downregulation of miR-1266 and miR-185 was demonstrated in PCa tissues and cell lines. The aim of the present
study was to investigate whether miR-1266 and miR-185 are involved in the regulation of B-cell lymphoma (BCL) 2
and BCL2L1, respectively, and whether transfection of PCa cell lines with miR-1266 and miR-185 mimics can alter
tumorigenic phenotypes. Methods: In order to investigate the regulation of BCL2 and BCL2L1 mRNA levels by
miR-1266 and miR-185, respectively, a luciferase reporter assay was used. Real-time PCR was also used to analyze
changes in the levels of BCL2 and BCL2L1 mRNAs in PCa cell lines following transfection with synthetic miR-1266
and miR-185. Cell apoptosis was determined by Annexin V protein expression analysis via flow cytometry. In addition
to the MTT assay, a cell proliferation assay was performed. Result: A luciferase assay confirmed that the BCL2 and
BCL2L1 genes may be targeted by miR-1266 and miR-185, respectively, through binding to their 3′UTR regions.
Transfection of PC3 and DU145 cells with miR-1266 and miR-185 induced apoptosis and reduced proliferation, which
also revealed an inverse correlation with BCL2 and BCL2L1 gene expression in the treated cells. Conclusion: Our
data suggests that miR-1266 and miR-185 may be novel candidates for further research in PCa treatment through the
anti-apoptotic pathway.

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