INTRODUCTION:Vascular endothelial growth factor (VEGF) regulates several cell functions including; proliferation, differentiation, permeability, vascular tone, and the production of vasoactive molecules.
The purpose of this study was to evaluate the potency of specific short-interfering RNA (siRNA) to suppress human VEGF expression by siRNA and investigate the effects of VEGF down-regulation on the cell proliferation and apoptosis of the human prostate cancer cell lines DU-145.
METHODS:Transfection was performed using X-tremeGENE siRNA transfection reagent. At different time intervals, transfected cells were harvested and total RNA was extracted for RT-PCR. The VEGF content in supernatants were measured by ELISA. Inhibition of cell growth by hVEGF-siRNA was measured by using cell proliferation ELISA BrdU assay. Apoptotic cells were evaluated by using annexin-V-FITC apoptotic detection method.
RESULTS:Transfection of hVEGF-siRNA resulted in statistically significant inhibition of hVEGF-mRNA that in turn caused a marked reduction in the expression of hVEGF. The cell growth was assessed every 24 h for 4 days after siRNA treatment resulted in a marked inhibition of cell proliferation as compared to scramble siRNA. The results of apoptosis showed that approximately 15 % of the cells treated with control-siRNA manifested evident apoptotic changes after 24 hpt, whereas DU-145 cells treated with hVEGF-siRNA significantly were positive, that is to say, 53 % at 72 hpt 23.9 ± 2.78 % (P < 0.001) and 13 ± 1.57 % at 96 hpt.
CONCLUSION: Our findings indicate that siRNA are effective in eliciting the RNAi pathway in cancerous cells and that specific siRNA efficiently down-regulate VEGF expression. They could decrease VEGF production and induce apoptosis, which may also be linked to the inhibition of cancerous cell proliferation. Therefore, it can be concluded that siRNA-mediated suppression of VEGF represents a powerful tool against prostate cancer cell proliferation. VEGF down-regulation exerts a direct anti-apoptotic function in the DU-145 cell lines and promises the development of drugs for cancer therapy.
Written by:
Deezagi A, Ansari-Majd S, Vaseli-Hagh N. Are you the author?
Department of Biochemistry, National Institute of Genetic Engineering and Biotechnology, Km. 17, Karaj-Tehran Freeway, Pajouhesh Blvd., P.O.Box 14155-6343, Tehran, Iran.
Reference: Clin Transl Oncol. 2012 Jul 24. Epub ahead of print.
doi: 10.1007/s12094-012-0868-1
PubMed Abstract
PMID: 22855159
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