摘要
目的:探讨锌指蛋白28(ZFP28)对肾透明细胞癌(ccRCC)细胞增殖、代谢、运动的影响及其机制.方法:采用免疫印迹和qPCR检测786-O和HK-2细胞中ZFP28的表达.CCK-8实验、集落形成实验、Edu实验、ATP合成实验、葡萄糖摄取实验、流式细胞术检测细胞的增殖及代谢能力;免疫印迹实验显示ZFP28对AKT/mTOR通路的影响;Transwell实验揭示ZFP28对细胞迁移与侵袭的影响.结果:ZFP28在ccRCC细胞786-O中与正常肾近端小管的上皮细胞系HK-2相比高表达(P＜0.05).通过siRNA导致的ZFP28下调抑制786-O细胞的生长与集落形成、ATP合成和葡萄糖摄取,并抑制786-O细胞的迁移及侵袭(P＜0.05).ZFP28下调抑制786-O细胞的AKT/mTOR通路(P＜0.05).结论:ZFP28通过AKT/mTOR轴抑制ccRCC细胞的生长、代谢和运动,可作为ccRCC治疗的潜在靶点.
Abstract
Objective To investigate the influence of ZFP28 on proliferation,metabolism,and motility of clear cell renal cell carcinoma(ccRCC)cells,along with its underlying mechanism.Methods Immunoblotting and qPCR were employed to assess the expression of ZFP28 in 786-O cells and HK-2 cells.Cell proliferation and metabolism capacity were determined using CCK-8 assay,colony formation,Edu,ATP synthesis assay,glucose uptake assay,and flow cytometry.The impact on the AKT/mTOR pathway was evaluated through immunoblotting,while Transwell assays were conducted to reveal effects on cell migration and invasion.Results ZFP28 was highly expressed in ccRCC cells compared with normal proximal renal tubule epithelial cell line HK-2(P＜0.05).Down-regulation of ZFP28 by siRNA inhibited the growth,colony formation,ATP synthesis and glucose uptake of 786-O cells,and inhibited the migration and invasion of 786-O cells(P＜0.05).ZFP28 down-regulated the AKT/mTOR pathway of 786-O cells in ccRCC cells(P＜0.05).Conclusion ZFP28 inhibits the growth,metabolism,and motility of clear cell renal cell carcinoma cells through the AKT/mTOR axis,suggesting its potential as a therapeutic target for ccRCC.
维普
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