【摘要】 目的采用反义寡核苷酸技术了解HSP70与细胞衰老之间的关系。方法设正义组、反义组,在转染后12,24 h,3,5 d,分别用流式细胞技术和免疫荧光技术检测HSP70蛋白水平、凋亡细胞比例和细胞周期。在转染后第2、4、6天,分别收集正义组、反义组和空白对照组细胞爬片进行SA-β-Gal染色。结果各时间点反义组蛋白表达均低于正义组(P<0.05),凋亡细胞比例均高于正义组(P<0.05)。随观察期延长,处于G1期的细胞数逐渐增多,但反义组均高于正义组(P<0.05)。相同时间点反义组SA-β-Gal染色阳性细胞(蓝染细胞)数均较正义组和空白对照组多,随观察期延长,各组蓝染细胞均增多。结论HSP70反义寡脱氧核苷酸下调MRC-5细胞HSP70的表达,使细胞出现G1期阻滞,促进细胞衰老。更多还原

【Abstract】 Objective To explore the relationship between heat shock protein 70(HSP70) and cell senescence by anti-sense oligodeoxynucleotides.Methods Flow cytometry and immunofluorescence were applied to measure the level of HSP70 protein,the proportion of apoptosis and cell cycle in plus sense and antisense group at 12 hours,1,3 and 5 days respectively after transfection.SA-β-Gal staining was used to detect the senescence cells at 2,4 and 6 days after transfection in plus sense,antisense and control group.Results At each time point,average fluorescence intensity was lower(P<0.05) and percentage of the apoptosis cells of anti-sense group was significantly higher than those of sense group(P<0.05).Cells arrested in the G1 phase increased with time,but the anti-sense group had significantly higher percentage than the sense group(P<0.05).SA-β-Gal staining showed that the number of positive cells in anti-sense group was more than those in sense or control group,and the positive cells increased with time in every group.Conclusions HSP70 antisense oligodeoxynucleotides can downregulate the HSP70 protein expression in the MRC-5 cell and induce the arrested cells in G1 phase to promote cell senescences.更多还原