Effects of lentiviral RNA interference-mediated downregulation of integrin-linked kinase on biological behaviors of human lens epithelial cells  被引量:2

Effects of lentiviral RNA interference-mediated downregulation of integrin-linked kinase on biological behaviors of human lens epithelial cells

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作  者:Yu-Ping Zheng Shao-Bo Zhang Feng Wang Hui Liu Wen Zhang Bin Song Zi-Yao Liu Lei Xiong Ya-Zhi Fan Ding-Ying Liao 

机构地区:[1]Department of Ophthalmology, the Second Affiliated Hospital, School of Medicine, Xi'an Jiaotong University [2]Department of Ophthalmology, Tangdu Hospital, the Fourth Military Medical University [3]Department of Otolaryngology, Shaanxi Provincial People's Hospital

出  处:《International Journal of Ophthalmology(English edition)》2016年第1期21-28,共8页国际眼科杂志(英文版)

基  金:Supported by the National Natural Science Foundation of China(No.81273605,No.30901655)

摘  要:AIM:To investigate the effects of lentivirus(LV)mediated integrin-linked kinase(ILK)RNA interference(RNAi)on biological behaviors of human lens epithelial cells(LECs).·METHODS:Human cataract LECs and immortalized human LEC line,human lens epithelial(HLE)B-3 cells were transfected by lentiviral vector expressing ILKspecific short hairpin RNA(sh RNA)and then stimulated by transforming growth factor-β(TGF-β),the silencing of ILK gene and protein was identified by reverse transcription-polymerase chain reaction(RT-PCR)and Western blot methods;biological behaviors including cell cycle and apoptosis,cell morphology,α-smooth muscle actin(SMA)stress fiber formation and cell migration were examined.·RESULTS:Remarkable decreases of ILK protein expression were detected in LECs carrying lentiviral ILK-sh RNA vector;flow cytometry revealed arresting of cell cycle progression through the G1/S transition and higher apoptosis rate in ILK-RNAi-LV transfected cells.Lessα-SMA stress fiber formation and migration was observed in ILK-RNAi-LV transfected LECs.·CONCLUSION:The present study demonstrated that ILK was an important regulator for LECs proliferation and migration.LV mediated ILK RNAi is an effective way todecrease ILK-regulated cell growth by arresting cell cycle progression and increasing cell apoptosis,as well as,to prevent cell migration by inhibiting TGF-βinducedα-SMA stress fiber formation.Thus,LV mediated ILK RNAi might be useful to prevent posterior capsular opacification.AIM:To investigate the effects of lentivirus(LV)mediated integrin-linked kinase(ILK)RNA interference(RNAi)on biological behaviors of human lens epithelial cells(LECs).·METHODS:Human cataract LECs and immortalized human LEC line,human lens epithelial(HLE)B-3 cells were transfected by lentiviral vector expressing ILKspecific short hairpin RNA(sh RNA)and then stimulated by transforming growth factor-β(TGF-β),the silencing of ILK gene and protein was identified by reverse transcription-polymerase chain reaction(RT-PCR)and Western blot methods;biological behaviors including cell cycle and apoptosis,cell morphology,α-smooth muscle actin(SMA)stress fiber formation and cell migration were examined.·RESULTS:Remarkable decreases of ILK protein expression were detected in LECs carrying lentiviral ILK-sh RNA vector;flow cytometry revealed arresting of cell cycle progression through the G1/S transition and higher apoptosis rate in ILK-RNAi-LV transfected cells.Lessα-SMA stress fiber formation and migration was observed in ILK-RNAi-LV transfected LECs.·CONCLUSION:The present study demonstrated that ILK was an important regulator for LECs proliferation and migration.LV mediated ILK RNAi is an effective way todecrease ILK-regulated cell growth by arresting cell cycle progression and increasing cell apoptosis,as well as,to prevent cell migration by inhibiting TGF-βinducedα-SMA stress fiber formation.Thus,LV mediated ILK RNAi might be useful to prevent posterior capsular opacification.

关 键 词:human lens epithelial cells integrin-linkedkinase RNA interference LENTIVIRUS posterior capsularopacification 

分 类 号:R776[医药卫生—眼科]

 

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