Effect Of TFPI Gene On The Proliferation Of Prostate Smooth Muscle Cells

Objective 1.To establish the method for isolating and culturing human primary prostate smooth muscle cells in vitro.2.To study the effects of TFPI gene on the proliferation of the human prostate smooth muscle cells.Methods The prostate tissues were isolated from BPH patients by surgery and the prostate SMOs were subsequently isolated and cultured by collagenase digestion methods.The isolated prostate SMOs were confirmed by immunohistochemical staining and MASSON staining. The passage 5 of the cells was used for the subsequent experiments.The cells were divided into two groups,control group and TFPI gene transfection groups.The cell numbers were counted on differrent days after the gene transfection.Reverse transcription polymerase chain reaction(RT-POR) was employed for the detection of the TFPI mRNA expression in the cells.Results 1.It was shown that the cultured prostate SMOs grew in a manner of peak and valley appearance.The prostate SMCs reached the logarithmic growth phase at 3 to 5 days after cell passing.2.The TFPI mRNA level in the cells transfected with the TFPI gene is 6 fold higher than that in the untreated cells and 2.5 fold higher than that of the cells transfected with the plRES DNA.3.The proliferation of the prostate smooth muscle cells was significantly inhibited by the TFPI gene beginning from the day 4 after the gene transfection.Conclusions 1.The collagenase digestion method is a suitable assay for the isolation of human primary prostate smooth muscle cells,which is able to provide sufficient number of cells of high activities for the gene transfection experiment.2 The expression of the TFPI gene in the cells was significantly elevated after the TFPI gene transfection.3.The proliferation of the cultured prostate smooth muscle cells was significantly inhibited by the TFPI gene,which suggested that TFPI might have a regulatory effect on the proliferation of the prostate SMC. 4.Further studies are needed for the understanding of the mechanism by which the TFPI gene regulate the proliferation of prostate SMCs.