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Blockade of insulin receptor substrate-1 inhibits biological behavior of choroidal endothelial cells

International Journal of Ophthalmology(English edition) 2019年第9期12卷 1386-1394页

作者： Yi-Yong Qian Hong-Ya Wu Gao-Qin Liu Chi Ren Pei-Rong Lu Xue-Guang Zhang Department of Ophthalmology the First Affiliated Hospital of Soochow UniversitySuzhou 215006Jiangsu ProvinceChina Department of Ophthalmology Shanghai Tenth People’s HospitalTongji University School of MedicineShanghai 200072China Jiangsu Key Laboratory of Clinical Immunology the First Affiliated Hospital of Soochow UniversitySuzhou 215006Jiangsu ProvinceChina

AIM: To investigate the effects of blockade of insulin receptor substrate-1(IRS-1) on the bio-function of tube formation of human choroidal endothelial cells(HCECs).METHODS: Quantitative reverse transcriptionpolymerase chain reaction(RT-PCR) and Western blot were performed to determine the expression level of IRS-1 and phospho-IRS-1 in HCECs. Tube formation of HCECs was analyzed using three dimensional in vitro Matrigel assay with or without IRS-1 blockage via IRS-1 inhibitor(GS-101) and vascular endothelial growth factor receptor 2(VEGFR2) inhibitor. In addition, cell counting kit(CCK)-8 and Transwell migration assay were exerted to analyze the effects of blockade of IRS-1 on the bio-function of proliferation and migration of HCECs, respectively. The apoptosis of HCECs was examined using flow cytometry(FCM).RESULTS: RT-PCR and Western blot revealed that IRS-1 phospho-IRS-1 were expressed in HCECs and the expression level was enhanced by stimulation of VEGF-A. The number of tube formation was decreased significantly in GS-101 treated groups compared to phosphate buffered saline(PBS) treated control groups. Furthermore, both cell proliferation and migration of HCECs were decreased in the presence of GS-101. FCM analysis showed that the apoptosis of HCECs was enhanced when the cells were treated with GS-101. Western blot also showed that the expression level of cleaved-caspase 3 in GS-101 treated group was higher than that in control ***: Blockade of IRS-1 can inhibit tube formation of HCECs through reducing cell proliferation and migration and promoting cell apoptosis.

关键词： insulin receptor substrate-1 choroidal endothelial cells neovascularization proliferation

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