The role of mechanical stretch and TGF-β2 in epithelial-mesenchymal transition of retinal pigment epithelial cells  

作　　者：Qian Cao Qu-Zhen Deji Ya-Jun Liu Wei Ye Wang-Dui Zhaba Qin Jiang Feng Yan 

机构地区：[1]Eye Hospital,Nanjing Medical University,Nanjing 210002,Jiangsu Province,China [2]The Fourth School of Clinical Medicine,Nanjing Medical University,Nanjing 210002,Jiangsu Province,China [3]Affiliated Jinling Hospital,Medical School of Nanjing University,Nanjing 210002,Jiangsu Province,China [4]Departm ent of Ophthalm ology,N anjing Drum Tower Hospital,The Affiliated Hospital of Nanjing University Medical School,Nanjing 210008,Jiangsu Province,China [5]School of Second Military Medical University,Department of Ophthalmology,Jinling Hospital,Nanjing 210002,Jiangsu Province,China

出　　处：《International Journal of Ophthalmology(English edition)》2019年第12期1832-1838,共7页国际眼科杂志（英文版）

基　　金：Supported by the National Natural Science Foundation of China (No.81600754)

摘　　要：AIM: To explore the effects and mechanisms of mechanical stress and transforming growth factor-beta2(TGF-β2) on epithelial-mesenchymal transition(EMT) in cultured human retinal pigment epithelial(RPE) cells. METHODS: Human RPE cells were inoculated on BioF ex 6-well plates and RPE cells received 0, 1, 2, 3, or 4 mild stretch injuries delivered 3h apart after 24h of culture. The device of mechanical stress parameters were set to sine wave, frequency 1 Hz, stretch strength 20%. For treatment with TGF-β2, when the inoculated RPE cells in 6-well plates were around 60% confluent, serum was reduced to 0 for 12h and recombinant human TGF-β2(0, 1, 5, 10 ng/mL)was added for 48h. α-SMA, Vimentin and N-Cadherin, fibronectin proteins expressions were detected by Western blotting, confocal cell immunofluorescence and quantitative real-time polymerase chain reaction(q RT-PCR). Then we detected the change of mi RNA-29b and ascertained the changes of phosphatidylinositol 3-kinase-serine threonine protein kinase(PI3K/Akt) pathway after RPE cells were stretched by the device of mechanical stress and induced by TGF-β2 by Western blotting, confocal cell immunofluorescence and qR T-PCR. RESULTS: Mechanical stress induce EMT and activate the PI3K/Akt pathway in ways that lead to the EMT process. TGF-β2 induce RPE cells EMT and in a certain range and TGF-β2 decrease the miR NA-29b expression in RPE cells, and the inhibitory effect is more obvious with the increase of TGF-β2 concentration. CONCLUSION: Our findings are crucial steps in determining the critical roles of the PI3K/Akt signaling pathway and mi RNA-29b in pathogenesis of proliferative vitreoretinopathy(PVR) which may be a potential target for preventing or treating PVR.● AIM: To explore the effects and mechanisms of mechanical stress and transforming growth factor-beta2(TGF-β2) on epithelial-mesenchymal transition(EMT) in cultured human retinal pigment epithelial(RPE) cells.● METHODS: Human RPE cells were inoculated on BioF ex 6-well plates and RPE cells received 0, 1, 2, 3, or 4 mild stretch injuries delivered 3h apart after 24h of culture. The device of mechanical stress parameters were set to sine wave, frequency 1 Hz, stretch strength 20%. For treatment with TGF-β2, when the inoculated RPE cells in 6-well plates were around 60% confluent, serum was reduced to 0 for 12h and recombinant human TGF-β2(0, 1, 5, 10 ng/mL)was added for 48h. α-SMA, Vimentin and N-Cadherin, fibronectin proteins expressions were detected by Western blotting, confocal cell immunofluorescence and quantitative real-time polymerase chain reaction(q RT-PCR). Then we detected the change of mi RNA-29b and ascertained the changes of phosphatidylinositol 3-kinase-serine threonine protein kinase(PI3K/Akt) pathway after RPE cells were stretched by the device of mechanical stress and induced by TGF-β2 by Western blotting, confocal cell immunofluorescence and qR T-PCR.● RESULTS: Mechanical stress induce EMT and activate the PI3K/Akt pathway in ways that lead to the EMT process. TGF-β2 induce RPE cells EMT and in a certain range and TGF-β2 decrease the miR NA-29b expression in RPE cells, and the inhibitory effect is more obvious with the increase of TGF-β2 concentration.● CONCLUSION: Our findings are crucial steps in determining the critical roles of the PI3K/Akt signaling pathway and mi RNA-29b in pathogenesis of proliferative vitreoretinopathy(PVR) which may be a potential target for preventing or treating PVR.

关 键 词：mechanical stress transforming growth factor-beta2 microRNA 29b epithelial-mesenchymal transition phosphatidylinositol 3-kinase-serine threonine protein kinase pathway proliferative vitreoretinopathy 

分 类 号：R73[医药卫生—肿瘤]