Objective To study the effect of a delivery system for administering bFGF-mRNA antisense oligodeoxynucleotide (AS-ODN) to prostatic stromal cells in vitro.Methods Human prostate stromal cells were confirmed to express a smooth muscle phenothype by morphological criteria and immunohistochemistry .Human prostatic stromal cells was transfected with a synthetic oligodeoxynucleotide complementary to the 5 ' end region of bFGF mRNA . Proliferation of human prostate stromal cells was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay . Semiquantative RT-PCR identified bFGF-mRNA in human prostate stromal cells .Results The majority of the cells stained positively for a -smooth muscle actin and vimentin indicates predominant smooth muscle and fibroblast phenotype.The strongest inhibitory effect was at 24 hours after the bFGF-mRNA AS-ODN transfected by liposome,the optimal concentration of bFGF mRNA antisense ODN was 0.8umol/L in thepresence of 1.0 ul of Oligofectamine Reagent , the proliferation of human prostate stromal cells was inhibited by 50.4% (p<0.05)with thelower expression of bFGF-mRNA (p<0.05). Without liposome,the bFGF-mRNA AS-ODN had no inhibitory effect to the prostate stromal cells in vitro even if the concentration of antisense ODN was 8mol/L. Conclusions Transfection mediated by liposome was efficient. bFGF may have a important role as a mediator of stromal expansion in benign prostatic hyperplasia(BPH). The results demonstrated the feasibility of bFGF-mRNA as a target for antisense gene therapy of human BPH and provide the theoretical foundation and experimental evidence for further study.
|