Adenovirus-mediated delivery of shRNA against bFGF mRNA suppresses growth of cultured human primary prostatic stromal cells

作者：Wang, Kai; Cheng, Linfeng; Liang, Yinghong; Liu, Donghui; Li, Kai; Wang, Ping^*

来源：Molecular Biology Reports, 2011, 38(2): 971-976.

DOI：10.1007/s11033-010-0191-2

• RNA interference
• Adenovirus
• Small hairpin RNA
• Basic fibroblast growth factor (bFGF)
• Human prostatic stromal cells (PrSCs)

Overexpression of basic fibroblast growth factor (bFGF) has been implicated in the pathogenesis of benign prostatic hyperplasia (BPH) and bFGF has been considered to be a promising therapy target for BPH. RNA interference (RNAi) based therapeutic approaches hold promise for the treatment of a variety of diseases. However, RNAi experiments have seldom been performed in human prostatic stromal cells (PrSCs). In the present study, we tranfected adenovirus type 5 vector mediated small hairpin RNA (shRNA) against human bFGF mRNA (Ad-sh-bFGF) to examine the proliferation and apoptosis effects on cultured human primary PrSCs. The gene-silencing effect of shRNA was evaluated by western blot. Cell proliferation was determined by MTT assays. Cell apoptosis was analyzed by flow cytometry and detection of caspase-3 activity. The effect of Ad-sh-bFGF on Bcl-2 gene expression was also examined. Adenovirus type 5 can efficiently delivered shRNA against bFGF into to PrSCs and the level of protein was depressed significantly in cells infected by Ad-sh-bFGF, approximately 50% lower than those cells infected by adenovirus-delivered nonsense shRNA (P < 0.01). Moreover, Ad-sh-bFGF is able to induce apoptosis and inhibit proliferation of cultured human primary PrSCs significantly (P < 0.01). Bcl-2 protein expression was markedly inhibited by transfection with Ad-sh-bFGF. In conclusion, our findings suggest that RNAi delivered via an adenovirus vector offers a prospect of improvement in treatment of BPH and bFGF is a potential target worth exploiting in BPH.

• 出版日期2011-2
• 单位中国医科大学