Overexpression of Stem Cell Protein, PIWIL2, Promotes the Invasive Characteristics of DU145 Prostate Cancer Cell Line

Publish Year: 1397
نوع سند: مقاله کنفرانسی
زبان: English
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NSCMRMED03_239

تاریخ نمایه سازی: 30 دی 1397

Abstract:

Background and Aim: Prostate cancer is the second cause of cancerdeath in the world. Invasion of malignant tumor cells via epithelial to-mesenchymal transition (EMT) is the main cause of death in cancerpatients. The stem cell protein, PIWIL2, is a member of the Argonautefamily, plays an important role in the EMT process by controlling andenhancing the invasive properties of cells. In this study, we aimed toexamine the effect of overexpression of piwil2 gene and its effects onEMT process in the prostate cancer cell line, DU145.Methods: DU145 cell line was cultured in the RPMI media containing10% fetal bovine serum (FBS) with 1% penicillin-streptomycin (Pen-Strep) suggested by ATCC. In order to provide DU145-PIWIL2 cell line,these cells were transfected by PCDNA3 vector expressing human piwil2gene driven by CMV promoter via electroporation method. To provideDU145-mock cells, PCDNA3 empty vector was transferred to the cellsvia electroporation. G418 was then applied to the transfected cells for4 weeks to establish stable cell lines before further cell and molecularanalysis. To investigate the effect of Piwil2 gene overexpression on cellgrowth, the doubling time method was performed to compare the cellsgrowth rate between DU145-PIWIL2 and DU145-mock cells. Moreover,the expression of EMT biomarkers was investigated in the two cell linesusing RT-PCR and Real-Time PCR.Results: The doubling time analysis profiled a significant increase inDU145-PIWIL2 cells growth rate compared to DU145-mock cells,exhibiting 2.31 times higher growth in the cells overexpressing PIWIL2.Quantitative real-time PCR indicated changes in some canonicalEMT biomarkers including E-cadherin, Vimentin, and Snail, with 0.77decreases in E-cadherin expression, as well as 1.36 and 2.01 increase inVimentin and Snail expressions respectively.Conclusion: In this study our data showed that overexpression of Piwil2gene promoted the growth rate of DU145 cells and induced the EMTprocess in these cells, hence enhancing the invasive potential of cancercells. Therefore, these data profiled piwil2 as an important biomarkerin assessing the malignant state of prostate cancer with application inprocedures related to cancer diagnostics and targeted therapies.

Authors

Bahar Hejazi Pishkenari

Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran

Shiva Irani

Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran

Hamid Reza Soleimanpour Lichaei

Department of Stem Cells and Regenerative Medicine, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran