缺氧抑制RABEP1基因表达对肾癌细胞786-O凋亡及增殖的影响

摘要

目的：探讨缺氧对RABEP1表达水平的影响及其表达情况对肾癌细胞786-O增殖及凋亡能力的影响。方法常氧及缺氧条件下培养人肾癌细胞786-O，荧光定量RT-PCR及蛋白印迹法检测RABEP1的表达情况；在缺氧条件下过表达RABEP1基因，MTT实验及Hoechst实验检测786-O细胞的增殖及凋亡情况。结果相对常氧培养的786-O细胞，缺氧可抑制细胞内RABEP1 mRNA及蛋白的表达水平，p值分别为0.0015及0.008；在第3、4及5天，相对786-OMOCK细胞，缺氧条件下786-OpcDNA-RABEP1细胞的增殖能力明显减弱，P值均<0.001；同样，相对786-OMOCK细胞，缺氧条件下786-OpcDNA-RABEP1细胞的凋亡明显增加。结论缺氧可抑制Rabaptin-5的表达，缺氧环境下过表达Rabaptin-5可抑制肾癌细胞的增殖并促进细胞凋亡。%Objective To explore the expression of RABEP1 in human renal cancer cell 786-O and investigate the influence of RABEP1 expression on proliferation and apoptosis under hypoxia. Methods The human renal cancer cell 786-O was cultured in normoxic and hypoxic condition. The mRNA and protein levels of RABEP1were detected by qRT-PCR and Western blot.The 786-O cell with ectopic expression of RABEP1 was cultured under hypoxia,and MTT and Hoechst tests were applied to investigate proliferation and apoptosis ability of 786-O cell. Results Compared to 786-O cell under normoxia,RABEP1 mRNA and protein expression level were down-regulated under hypoxia,P=0.0015 and 0.008,respectively. Compared to 786-OMOCK under hypoxia from day 3 to day 5,proliferation ability of 786-OpcDNA-RABEP1 cell was inhibited under hypoxia,all the p values were less than 0.001. Then,Compared to 786-OMOCK under hypoxia,apoptosis ability of 786-OpcDNA-RABEP1 cell was enhanced under hypoxia. Conclusions Hypoxia could inhibit RABEP1 expression,and ectopic expression of RABEP1 could suppress proliferation and enhance apoptosis of human renal cancer 786-O under hypoxia.