Adenovirus-mediated delivery of shRNA against bFGF mRNA suppresses growth of cultured human primary prostatic stromal cells.
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 transfected 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.
Wang
Kai
K
Department of Urology, The Fourth Affiliated Hospital of China Medical University, 4 East Chongshan Road, Shenyang, 110032, People's Republic of China.
Cheng
Linfeng
L
Liang
Yinghong
Y
Liu
Donghui
D
Li
Kai
K
Wang
Ping
P
D017209:Apoptosis; D001483:Base Sequence; D053148:Caspase 3 / Q000378:metabolism; D049109:Cell Proliferation; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism*; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D011467:Prostate / Q000378:metabolism*; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics*; D017154:Stromal Cells / Q000378:metabolism*; D013778:Tetrazolium Salts / Q000494:pharmacology; D013844:Thiazoles / Q000494:pharmacology
D012333:RNA, Messenger; D034741:RNA, Small Interfering; D013778:Tetrazolium Salts; D013844:Thiazoles; D016222:Fibroblast Growth Factor 2; D053148:Caspase 3; C022616:thiazolyl blue