Background:Estrogen is involved in the pathophysiological process of benign prostatic hyperplasia(BPH),in which epithelial-mesenchymal transition(EMT)plays an important role.Upregulation of aquaporin(AQP)5,which is di...Background:Estrogen is involved in the pathophysiological process of benign prostatic hyperplasia(BPH),in which epithelial-mesenchymal transition(EMT)plays an important role.Upregulation of aquaporin(AQP)5,which is directly activated by estrogen,has been reported to promote EMT in multiple cells.This study aimed to examine the effects of AQP5 on estrogen-induced EMT in the prostate.Methods:Normal prostate(NP)tissue samples without any histopathological changes and BPH tissue samples with pathologically confirmed hyperplasia were obtained.An EMT cell model was subsequently established by adding estradiol(E2)to RWPE-1 cells,after which AQP5 knockdown was performed.Tissue morphological and immunohistochemical features were examined using hematoxylin-eosin and immunohistochemical staining.Western blot analysis was performed to determine the expression of AQPs,estrogen receptors,and EMT-related proteins.Cell proliferation was assessed and supernatants were collected for enzyme-linked immunosorbent assay to determine transforming growth factor-β1(TGF-β1)concentrations.Immunofluorescence staining was performed to assess protein expressions in RWPE-1 cells.Results:BPH tissues exhibited greater EMT(TGF-β1:1.362±0.196 vs.0.107±0.067,P=0.003;vimentin:1.581±0.508 vs.0.221±0.047,P<0.001;E-cadherin:0.197±0.188 vs.1.344±0.088,P<0.001),higher AQP5(1.268±0.136 vs.0.227±0.055,P<0.001)and estrogen receptor(ER)α(1.250±0.117 vs.0.329±0.134,P<0.001)expression but lower ERβ(0.271±0.184 vs.1.564±0.130,P<0.001)expression than NP tissues.E2-stimulated cells had higher AQP5 expression(1.298±0.058 vs.1.085±0.104,P=0.049),increased cell proliferation(1.510±0.089 vs.1.000±0.038,P<0.001),and EMT(TGF-β1 concentration:0.352±0.021 ng/mL vs.0.125±0.014 ng/mL,P<0.001;vimentin:1.641±0.120 vs.0.188±0.020,P=0.002;E-cadherin:0.075±0.030 vs.0.843±0.046,P<0.001)than controls.E2-stimulated cells with AQP5 knockdown exhibited decreased EMT(TGF-β1 concentration:0.223±0.041 ng/mL vs.0.352±0.021 ng/mL,P=0.016;vimentin:0.675±0.056 vs.1.641±0.120,P=0.001;E-cadherin:0.159±0.037 vs.0.075±0.030,P=0.040)than E2-stimulated cells with non-related small interfering RNA(siRNA).Conclusion:Our findings suggest that estrogen induces BPH possibly by promoting AQP5 expression.Hence,AQP5 might be a novel target for modulating EMT in prostate epithelial cells.展开更多
Resveratrol is a dietary polyphenol espoused to have chemopreventive activity against a variety of human cancer types. We first reported that resveratrol significantly decreases the proliferation of both androgen-depe...Resveratrol is a dietary polyphenol espoused to have chemopreventive activity against a variety of human cancer types. We first reported that resveratrol significantly decreases the proliferation of both androgen-dependent and hormone-refractory prostate cancer cells. However, the effects of resveratrol in normal prostate epithelial and stromal cells, particularly with regard to its uptake, subcellular distribution and intracellular targets, have not been investigated. To advance the knowledge on accessibility and cellular disposition of resveratrol in prostate cells, [3H] resveratrol, fractionation of cell extracts into subcellular compartments, Western blot analysis, resveratrol affinity column chromatography and flow cytometry were used to study the uptake and intracellular distribution of resveratrol in normally cultured prostate stromal （PrSCs） and epithelial cells （PrECs）. Pretreatment of both PrSCs and PrECs for 2 days with resveratrol modulated its uptake and selectively increased its distribution to the membrane and organelle compartments. Resveratrol affinity column chromatography studies showed differential expression of a previously identified resveratrol-targeting protein, quinone reductase 2 （QR2）, in PrSCs and PrECs. Flow cytometric analysis comparing resveratrol-treated and untreated PrSCs showed a large decrease in G1-phase and a concomitant increase in S and G2/M-phases of the cell cycle. These results suggest that resveratrol suppresses PrSC proliferation by affecting cell cycle phase distribution, which may involve the participation by QR2.展开更多
基金This work was supported by grants from the National Key Research and Development Program of China(No.SQ2017YFSF090096)the National Natural Science Foundation of China(Nos.81974098,81770756,81974099,and 81300627)+1 种基金a Special Supportive Program for Organ Transplantation by COTDF(No.2019JYJH08)the Research Funding of Sichuan Health and Family Planning Commission(No.18PJ453)。
文摘Background:Estrogen is involved in the pathophysiological process of benign prostatic hyperplasia(BPH),in which epithelial-mesenchymal transition(EMT)plays an important role.Upregulation of aquaporin(AQP)5,which is directly activated by estrogen,has been reported to promote EMT in multiple cells.This study aimed to examine the effects of AQP5 on estrogen-induced EMT in the prostate.Methods:Normal prostate(NP)tissue samples without any histopathological changes and BPH tissue samples with pathologically confirmed hyperplasia were obtained.An EMT cell model was subsequently established by adding estradiol(E2)to RWPE-1 cells,after which AQP5 knockdown was performed.Tissue morphological and immunohistochemical features were examined using hematoxylin-eosin and immunohistochemical staining.Western blot analysis was performed to determine the expression of AQPs,estrogen receptors,and EMT-related proteins.Cell proliferation was assessed and supernatants were collected for enzyme-linked immunosorbent assay to determine transforming growth factor-β1(TGF-β1)concentrations.Immunofluorescence staining was performed to assess protein expressions in RWPE-1 cells.Results:BPH tissues exhibited greater EMT(TGF-β1:1.362±0.196 vs.0.107±0.067,P=0.003;vimentin:1.581±0.508 vs.0.221±0.047,P<0.001;E-cadherin:0.197±0.188 vs.1.344±0.088,P<0.001),higher AQP5(1.268±0.136 vs.0.227±0.055,P<0.001)and estrogen receptor(ER)α(1.250±0.117 vs.0.329±0.134,P<0.001)expression but lower ERβ(0.271±0.184 vs.1.564±0.130,P<0.001)expression than NP tissues.E2-stimulated cells had higher AQP5 expression(1.298±0.058 vs.1.085±0.104,P=0.049),increased cell proliferation(1.510±0.089 vs.1.000±0.038,P<0.001),and EMT(TGF-β1 concentration:0.352±0.021 ng/mL vs.0.125±0.014 ng/mL,P<0.001;vimentin:1.641±0.120 vs.0.188±0.020,P=0.002;E-cadherin:0.075±0.030 vs.0.843±0.046,P<0.001)than controls.E2-stimulated cells with AQP5 knockdown exhibited decreased EMT(TGF-β1 concentration:0.223±0.041 ng/mL vs.0.352±0.021 ng/mL,P=0.016;vimentin:0.675±0.056 vs.1.641±0.120,P=0.001;E-cadherin:0.159±0.037 vs.0.075±0.030,P=0.040)than E2-stimulated cells with non-related small interfering RNA(siRNA).Conclusion:Our findings suggest that estrogen induces BPH possibly by promoting AQP5 expression.Hence,AQP5 might be a novel target for modulating EMT in prostate epithelial cells.
文摘Resveratrol is a dietary polyphenol espoused to have chemopreventive activity against a variety of human cancer types. We first reported that resveratrol significantly decreases the proliferation of both androgen-dependent and hormone-refractory prostate cancer cells. However, the effects of resveratrol in normal prostate epithelial and stromal cells, particularly with regard to its uptake, subcellular distribution and intracellular targets, have not been investigated. To advance the knowledge on accessibility and cellular disposition of resveratrol in prostate cells, [3H] resveratrol, fractionation of cell extracts into subcellular compartments, Western blot analysis, resveratrol affinity column chromatography and flow cytometry were used to study the uptake and intracellular distribution of resveratrol in normally cultured prostate stromal （PrSCs） and epithelial cells （PrECs）. Pretreatment of both PrSCs and PrECs for 2 days with resveratrol modulated its uptake and selectively increased its distribution to the membrane and organelle compartments. Resveratrol affinity column chromatography studies showed differential expression of a previously identified resveratrol-targeting protein, quinone reductase 2 （QR2）, in PrSCs and PrECs. Flow cytometric analysis comparing resveratrol-treated and untreated PrSCs showed a large decrease in G1-phase and a concomitant increase in S and G2/M-phases of the cell cycle. These results suggest that resveratrol suppresses PrSC proliferation by affecting cell cycle phase distribution, which may involve the participation by QR2.
