胰岛素对人肝癌HepG2细胞体外诱导人脐静脉内皮细胞血管形成能力的影响

摘要

Objective:To investigate the effects of different concentrations of insulin on the ability of hepatocellular carcinoma cell line (HepG2) in inducing human umbilical vein endothelial cells(HUVECs) angiogenesis and its possible mechanism.Methods:HepG2 was pre-cultured using complete medium with different concentrations of insulin to make conditioned medium.The effect of different conditioned medium on the migration and invasion ability of HUVECs was detected using transwell ventricles with matrigel matrix medium.The effect of different conditioned medium on the proliferation ability of HUVECs was detected by CCK-8 and EdU cell proliferation assay.Tube experiments in endothelial cells were used to detect the effect of different conditioned medium on the ability of HUVECs in tube formation.In the meanwhile,RT-PCR was used to detect the effect of different concentrations of insulin on the HepG2 cells in VEGF121,VEGF165 and COX-2 transcription level.Results:In a certain concentration range,the effect of the HepG2 cells on the migration and proliferafion ability and the ability of HUVECs in tube formation was positively associated with the concentration of insulin,and the effect of the HepG2 cells in the VEGF121,VEGF165 and COX-2 transcription level was positively associated with the concentration of insulin.Conclusion:In a certain concentration range,insulin can promote the ability of HepG2 cells in inducing HUVFCs angiogenesis by increasing the expression level of VEGF121,VEGF165,COX-2.%目的:探讨胰岛素对人肝癌细胞系HepG2体外诱导人脐静脉内皮细胞(HUVECs)血管形成能力的影响及其可能机制.方法:用含不同浓度胰岛素的完全培养基预培养肝癌细胞系HepG2制备条件培养基;应用预铺Matrigel基质胶的Transwell小室检测不同组条件培养基对HUVECs迁移能力的影响;运用CCK-8方法及EdU细胞增殖实验检测不同组条件培养基对HUVECs增殖能力的影响;运用内皮细胞成管实验检测不同组条件培养基对HUVECs成血管能力的影响;同时应用RT-PCR检测不同胰岛素浓度培养的HepG2细胞中血管内皮生长因子(VEGF) 121、VEGF165、环氧化酶2(COX-2)的转录水平.结果:在一定浓度范围内,HepG2细胞对HUVECs的侵袭迁移能力、增殖能力的影响,对HUVECs的成血管能力的影响,对HepG2细胞VEGF 121、VEGF165、COX-2转录水平的影响,均分别与胰岛素浓度呈正相关.结论:在一定浓度范围内,胰岛素可能通过上调HepG2细胞中VEGF121、VEGF165、COX-2的表达水平促进HepG2细胞诱导HUVECs血管形成能力.