Cone-rod homeobox transcriptionally activates TCF7 to promote the proliferation of retinal pigment epithelial and retinoblastoma cells in vitro  

作　　者：Na Zhao Ying-Ying Li Jia-Man Xu Mu-Yao Yang Yun-Zhe Li Thomas Chuen Lam Lei Zhou Qi-Hu Tong Jun-Tao Zhang Sheng-Zhan Wang Xin-Xin Hu Yu-Fei Wu Qin-Kang Lu Ting-Yuan Lang 

机构地区：[1]Ophthalmology Center,the Affiliated People’s Hospital of Ningbo University,Ningbo 315040,Zhejiang Province,China [2]Wenzhou Medical University,Wenzhou 325035,Zhejiang Province,China [3]Reproductive Medicine Center,the First Affiliated Hospital of Chongqing Medical University,Chongqing 400016,China [4]College of Bioengineering,Chongqing University,Chongqing 400044,China [5]Department of Gynaecology and Obstetrics,the First Affiliated Hospital of Chongqing Medical University/Department of Gynecology,The First Branch of The First Affiliated Hospital of Chongqing Medical University,Chongqing,400042 [6]State Key Laboratory of Maternal and Fetal Medicine of Chongqing,Chongqing Medical University,Chongqing 400016,China [7]Centre for Myopia Research,School of Optometry,The Hong Kong Polytechnic University,Hong Kong 999077,China [8]Centre for Eye and Vision Research(CEVR),17W Hong Kong Science Park,Hong Kong 999077,China [9]School of Optometry/Department of Applied Biology and Chemical Technology/Research Centre for SHARP Vision(RCSV)/The Hong Kong Polytechnic University,Hong Kong 999077,China

出　　处：《International Journal of Ophthalmology(English edition)》2024年第11期1995-2006,共12页国际眼科杂志（英文版）

基　　金：Supported by grants from the Zhejiang Medicine and Health Science and Technology Project(No.2018KY748);Ningbo Natural Science Foundation(No.2019A610352);Ningbo Major Scientific and Technological Research and“Unveiling and Commanding”Project(No.2021Z054);Chongqing Science&Technology Commission(No.CSTB2022NSCQ-MSX1413);Ningbo Clinical Research Center for Ophthalmology(No.2022L003);Ningbo Key Laboratory for Neuroretinopathy Medical Research,and the Project of NINGBO Leading Medical&Health Discipline(No.2016-S05).

摘　　要：AIM:To investigate the proliferation regulatory effect of cone-rod homeobox(CRX)in retinal pigment epithelium(RPE)and retinoblastoma(RB)cells to explore the potential application and side effect(oncogenic potential)of CRXbased gene therapy in RPE-based retinopathies.METHODS:Adult human retinal pigment epithelial(ARPE)-19 and human retinal pigment epithelial(RPE)-1 cells and Y79 RB cell were used in the study.Genetic manipulation was performed by lentivirus-based technology.The cell proliferation was determined by a CellTiter-Glo Reagent.The mRNA and protein levels were determined by quantitative real-time polymerase chain reaction(qPCR)and Western blot assay.The transcriptional activity of the promoter was determined by luciferase reporter gene assay.The bindings between CRX and transcription factor 7(TCF7)promoter as well as TCF7 and the promoters of TCF7 target genes were examined by chromatin immunoprecipitation(ChIP)assay.The transcription of the TCF7 was determined by a modified nuclear run-on assay.RESULTS:CRX overexpression and knockdown significantly increased(n=3,P<0.05 in all the cells)and decreased(n=3,P<0.01 in all the cells)the proliferation of RPE and RB cells.CRX overexpression and knockdown significantly increased and deceased the mRNA levels of Wnt signaling target genes[including MYC proto-oncogene(MYC),JUN,FOS like 1(FOSL1),CCND1,cyclin D2(CCND2),cyclin D3(CCND3),cellular communication network factor 4(CCN4),peroxisome proliferator activated receptor delta(PPARD),and matrix metallopeptidase 7(MMP7)]and the luciferase activity driven by the Wnt signaling transcription factor(TCF7).TCF7 overexpression and knockdown significantly increased and decreased the proliferation of RPE and RB cells and depletion of TCF7 significantly abolished the stimulatory effect of CRX on the proliferation of RPE and RB cells.CRX overexpression and knockdown significantly increased and decreased the mRNA level of TCF7 and the promoter of TCF7 was significantly immunoprecipitated by CRX antibody.CONCLUSION:CRX transcri

关 键 词：retinal pigment epithelial cell RETINOBLASTOMA cone-rod homeobox transcription factor 7 regenerative medicine tumorigenic potential 

分 类 号：R774.1[医药卫生—眼科]