良性前列腺增生间质细胞通过分泌TGF-β1破坏前列腺腺上皮屏障OA

Objective:To investigate the effect of stromal cells on the prostatic luminal epithelial barrier in be-nign prostatic hyperplasia(BPH)and its potential mechanism.Methods:The human normal prostatic stromal im-mortalized cell line WPMY-1 and benign prostatic epithelial cell line BHPrE-1 were selected.BHPrE-1 cells were di-vided into a control group and treatment groups.The control group was cultured with BHPrE-1 medium,while the treatment groups were cultured with BHPrE-1 medium containing different proportions of WPMY-1 conditioned me-dium,or BHPrE-1 medium or WPMY-1 conditioned medium with or without transforming growth factor-β1(TGF-β1)antibody.Specifically,the treatment groups included a 50％WPMY-1 conditioned medium group(cultured with 50％WPMY-1 conditioned medium+50％BHPrE-1 cell medium)and a 100％WPMY-1 conditioned medium group(cultured with 100％WPMY-1 conditioned medium).Other treatment groups were set as follows:a WPMY-1 condi-tioned medium group(cultured with 100％WPMY-1 conditioned medium),a TGF-β1 antibody group(cultured with BHPrE-1 cell medium containing 1 μg/ml TGF-β1 antibody),and a WPMY-1 conditioned medium+TGF-β1 antibody group(cultured with 100％WPMY-1 conditioned medium containing 1 μg/ml TGF-β1 antibody).Transepithelial electrical resistance(TER)meter and FITC-dextran Transwell permeability assay were used to detect TER values and fluorescence values in each group.RT-qPCR and Western blot were applied to measure the mRNA and protein expressions of E-Cadherin and Claudin-1 in cells of each group.ELISA was used to detect the concentration of TGF-β1 in the culture medium.Results:Compared with the control group,the TER values in the 50％and 100％WPMY-1 con-ditioned medium groups were decreased sequentially,and the fluorescence values were increased sequentially(all P＜0.05).Compared with the control group,there were no significant differences in the mRNA expression levels of E-Cadherin in BHPrE-1 cells between the 50％and 100％WPMY-1 conditioned medium groups(all P＞0.05),whereas the protein expression of E-Cadherin and the mRNA and protein expressions of Claudin-1 were decreased se-quentially(all P＜0.05).Compared with BHPrE-1 cell medium,the concentration of TGF-β1 in WPMY-1 condi-tioned medium was increased(P＜0.05).Compared with the control group,the TGF-β1 antibody group,and the WPMY-1 conditioned medium+TGF-β1 antibody group,the TER value was decreased and the fluorescence value was increased in the WPMY-1 conditioned medium group(all P＜0.05).There were no significant differences in TER values and fluorescence values among the control group,the TGF-β1 antibody group,and the WPMY-1 condi-tioned medium+TGF-β1 antibody group(all P＞0.05).Compared with the control group,the TGF-β1 antibody group,and the WPMY-1 conditioned medium+TGF-β1 antibody group,the protein expression of E-Cadherin and the mRNA and protein expressions of Claudin-1 were decreased in the WPMY-1 conditioned medium group(all P＜0.05).There were no significant differences in the mRNA and protein expressions of E-Cadherin and Claudin-1 among the control group,the TGF-β1 antibody group,and the WPMY-1 conditioned medium+TGF-β1 antibody group(all P＞0.05).Conclusion:Stromal cells in BPH downregulate the expressions of E-Cadherin and Claudin-1 in epithelial cells by secreting TGF-β1,thereby disrupting the prostatic luminal epithelial barrier.