HAI-1对肾细胞癌细胞自噬及凋亡调控的初步研究  

Effect of HAI-1 on Autophagy and Apoptosis in Renal Cell Cancer Cells

作　　者：张靖华[1] 吴秋杰[1] 刘金瑞[1] 王鹏鸽 孙夕然 王长安[1] ZHANG Jing-hua;WU Qiu-jie;LIU Jin-rui;WANG Peng-ge;SUN Xi-ran;WANG Chang-an(Zhengzhou Seventh People’s Hospital,Zhengzhou 450017,China)

机构地区：[1]河南省郑州市第七人民医院,河南郑州450017

出　　处：《中国肿瘤》2020年第9期715-720,共6页China Cancer

摘　　要：[目的]研究肝细胞生长因子激活因子1型抑制因子(hepatocyte growth factor activator inhibitor type 1,HAI-1)对肾细胞癌细胞自噬和凋亡的影响,探讨其发挥作用的可能机制。[方法]QRTPCR检测HAI-1在肾细胞癌组织和细胞系中的表达水平,选择低表达的肾细胞癌细胞系进行HAI-1过表达质粒的转染,分为对照组(NC组)和过表达质粒组(OE-HAI-1组)。QRT-PCR检测NC组和OE-HAI-1组细胞中HAI-1的表达水平;MTS实验检测NC组和OE-HAI-1组细胞的增殖率;Western blot法检测HAI-1对肾细胞癌细胞中自噬相关蛋白表达水平的影响;流式细胞仪检测NC组和OEHAI-1组细胞凋亡率;Western blot法检测NC组和OE-HAI-1组细胞中caspase3、Bax和Bcl-2蛋白的表达水平。[结果]QRT-PCR结果显示HAI-1在肾细胞癌组织中的表达低于其在癌旁组织中的表达,HAI-1在肾细胞癌细胞系ACHN、786-O和A-498中的表达低于其在肾小管上皮细胞HK-2中的表达(P<0.05)。选择HAI-1表达最低的肾细胞癌细胞系786-O转染HAI-1过表达质粒,qRT-PCR结果显示OE-HAI-1组细胞中HAI-1的表达增加(P<0.05);与NC组相比,OE-HAI-1组786-O细胞增殖能力下降。与NC组比较,OE-HAI-1组和加入自噬激活剂Rapamycin的OE-HAI-1+Rapamycin组细胞中Beclin1和LC3-Ⅱ蛋白表达降低,LC3-Ⅰ蛋白表达增加。与OE-HAI-1组相比,OE-HAI-1+Rapamycin组Beclin1和LC3-Ⅱ蛋白表达增加,LC3-Ⅰ蛋白表达降低。与NC组相比,OE-HAI组细胞凋亡增加,细胞中caspase3和Bax蛋白表达增加,Bcl-2蛋白的表达降低。[结论]HAI-1在肾细胞癌组织和细胞系中表达降低,并促进肾细胞癌细胞自噬和凋亡的发生。HAI-1可能是治疗肾细胞癌的候选分子靶标。[Purpose]To investigate the effect of hepatocyte growth factor activator inhibitor type 1(HAI-1)on autophagy and apoptosis in renal cell carcinoma cells,and its possible mechanisms.[Methods]The expression levels of HAI-1 were detected with qRT-PCR in renal cell carcinoma tissues and cell lines.The HAI-1 plasmids(OE-HAI-1 group)or blank plasmids(NC group)were transfected in low-expressing renal cell carcinoma cells.MTS was used to detect the cell proliferation rate;Western blot method was used to detect the effect of HAI-1 on autophagy-related proteins expression in renal cell carcinoma cells;flow cytometry was used to detect apoptosis rates in NC group and OE-HAI-1 group.The expressions of caspase3,Bax and Bcl-2 proteins were detected with Western blot.[Results]The qRT-PCR results showed that the expression of HAI-1 in renal cell carcinoma tissues was lower than that in cancer adjacent tissues,and the expression of HAI-1 in renal cell carcinoma cell lines ACHN,786-O,and A-498 was lower than that in renal tubular epithelial HK-2 cells(P<0.05).The renal cell carcinoma cell line 786-O with the lowest HAI-1 expression was selected to transfect the HAI-1 overexpression plasmid.The qRT-PCR results showed that HAI-1 expression was increased in the OE-HAI-1 cells(P<0.05).Compared with the NC group,the expressions of Beclin1 and LC3-Ⅱproteins were decreased,and the expression of LC3-Ⅰprotein was increased in the OE-HAI-1 cells and OE-HAI-1+Rapamycin group.Compared with the OEHAI-1 group,the expressions of Beclin1 and LC3-Ⅱproteins were increased and the expression of LC3-Ⅰprotein was decreased in the OE-HAI-1+Rapamycin group.Compared with the NC group,the apoptosis of the OE-HAI group was increased,the expression of caspase3 and Bax protein were increased,and the expression of Bcl-2 protein was decreased.[Conclusion]HAI-1 expression is downregulated in renal cell carcinoma tissues and cell lines,resulting in the autophagy and apoptosis in renal cell carcinoma cells.HAI-1 may be a candidate molecular target for the

关 键 词：肾细胞癌 肝细胞生长因子激活因子1型抑制因子 自噬 凋亡 

分 类 号：R737.11[医药卫生—肿瘤]