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

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Abstract

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.

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Authors and Affiliations

  1. Department of Urology, The Fourth Affiliated Hospital of China Medical University, 4 East Chongshan Road, Shenyang, 110032, People’s Republic of China

    Kai Wang, Linfeng Cheng & Ping Wang

  2. Department of Pathology, Jinqiu Hospital of Liaoning Province, Shenyang, 110015, People’s Republic of China

    Yinghong Liang

  3. Department of Urology, Jinqiu Hospital of Liaoning Province, Shenyang, 110015, People’s Republic of China

    Donghui Liu

  4. Department of Oncology, The First Affiliated Hospital of China Medical University, Shenyang, 110001, People’s Republic of China

    Kai Li

Authors
  1. Kai Wang
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  2. Linfeng Cheng
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  3. Yinghong Liang
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  4. Donghui Liu
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  5. Kai Li
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Corresponding author

Correspondence to Ping Wang.

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Supplementary Fig. 1

The sequencing oscillogram of adenovirus vector (type 5, ΔE1/E3) delivery of shRNA targeting bFGF. (TIFF 297 kb)

Supplementary Fig. 2a

Primary cultured PrPCs were infected with Ad-EGFP for 48 h and observed through a fluorescence microscope: a Light field (Original magnification × 200) (TIFF 2461 kb)

Supplementary Fig. 2b

Primary cultured PrPCs were infected with Ad-EGFP for 48 h and observed through a fluorescence microscope: b Fluorescence field (Original magnification × 200) (TIFF 2459 kb)

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Wang, K., Cheng, L., Liang, Y. et al. Adenovirus-mediated delivery of shRNA against bFGF mRNA suppresses growth of cultured human primary prostatic stromal cells. Mol Biol Rep 38, 971–976 (2011). https://doi.org/10.1007/s11033-010-0191-2

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  • DOI: https://doi.org/10.1007/s11033-010-0191-2

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