1. Department of Biology, Science and Research branch, Islamic Azad University 2. Department of Biology, Science and Research branch, Islamic Azad University 3. Department of Stem Cells and Regenerative Medicine, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran
Prostate cancer is the most common cause of malignancy in men. piwil2 as a stem cell protein and a member of argonaute gene family is highly expressed in many cancers. the most important causes of death in prostate cancer is metastasis which is caused by the invasive characteristics of prostate cancer cells under the control of the mediators of the epithelial to mesenchymal transition phenomenon or emt.
in this study, the effects of piwil2 overexpression on the invasive and proliferation characteristics of the pc3 cell line of prostate cancer was investigated.
Pc3 cell line was cultured in the rpmi media containing 10% fetal bovine serum (fbs) with 1% penicillin streptomycin (pen/strep) suggested by atcc. in order to provide pc3-piwil2 cell line, these cells were transfected by pcdh vector expressing human piwil2 gene driven by cmv promoter via electroporation method. puromycin was then applied to the transfected cells for 10 days to establish stable cell lines before further cell and molecular analysis. to investigate the effect of piwil2 gene overexpression on cell growth, doubling time method were performed and real time pcr methods was the undertaken to investigate the emt features between pc3-piwil2 and control pc3 cells.
The doubling time analysis showed a significant increase in pc3-piwil2 cells growth rate compared to pc3 control cells, exhibiting 1.14 times higher growth in the cells overexpressing piwil2 (p<0.05). changes in the expression level of ecadherin and vimentin as two main emt biomarkers showed that piwil2 overexpression influenced on emt.
In this study our data showed that piwil2 promotes the proliferative capacity of prostate cancer cells as well as influencing on the invasive characteristics of these cells, hence underpinning the importance of piwil2 gene as a valuable biomarker for the purpose of cancer diagnosis and therapy.