【摘要】 目的:研究HDAC6siRNA对食管鳞癌EC9706细胞中HDAC6的干扰效果,以及HDAC6表达下调对食管鳞癌细胞增殖能力、细胞周期的影响。方法:将食管鳞癌EC9706细胞分为未处理组、对照siRNA组和HDAC6siRNA组3组,后2组分别转染对照siRNA和HDAC6siRNA。采用半定量反转录-聚合酶链反应、蛋白质印迹及免疫细胞化学检测各组食管鳞癌EC9706细胞中HDAC6siRNA对HDAC6的干扰效果;采用CCK-8试剂盒检测各组细胞增殖能力的变化;运用流式细胞术检测各组细胞的细胞周期的变化。结果:HDAC6siRNA组的HDAC6mRNA表达的相对水平为0.044,HDAC6蛋白表达相对水平为0.114,与未处理组(0.951、0.924)和对照组(0.947、0.905)相比,HDAC6mR-NA和蛋白的表达均显著下调,t值分别为2.24和2.52,P<0.05;细胞增殖明显受抑制,且G0/G1期的比率为(68.55±2.01)%,显著高于未处理组(45.64±1.26)%和对照siRNA组(46.23±1.39)%,S期的比率(25.93±0.73)%显著低于未处理组(33.13±1.02)%和对照siRNA组(32.98±0.91)%,差异均有统计学意义,P<0.05。结论:HDAC6siRNA能有效下调食管鳞癌EC9706细胞中HDAC6mRNA和蛋白的表达;HDAC6表达下调能明显抑制食管鳞癌EC9706细胞的增殖并能诱导细胞周期静止在G0/G1期。更多还原

【Abstract】 OBJECTIVE: To investigate the expression of HDAC6 siRNA on HDAC6 mRNA and proteins,and study the effect of downregulation of HDAC6 expression on cell proliferation and cell cycle.METHODS: The experiment was divided into three groups,including untreated group,control siRNA group and HDAC6 siRNA group.HDAC siRNA and control siRNA were transfected into ESCC cell line EC9706 cells in HDAC6 siRNA group and control siRNA group,respectively.To investigate the expressions of HDAC6 mRNA and protein and verify the effect of interference by semi-quantitative RT-PCR and Western blotting as well as immunocytochemistry methods.To study the effect of downregulation of HDAC6 expression on cell proliferation by CCK-8 kit.To detecte the effect on cell cycle by Flow cytometry.RESULTS: The HDAC6 mRNA relstive expression level in HDAC6 siRNA group was 0.044,the HDAC6 protein relstive expression level was 0.114,compared with untreated group(0.951,0.924) and control siRNA group(0.947,0.905),HDAC6 mRNA and protein were significantly downregulated after transfection with HDAC6 siRNA(t value was 2.24 and 2.52);the cell proliferation was markedly inhibited.The results of cell cycle showed that percentage of cell number in G0/G1 phase in HDAC6 siRNA group(68.55±2.01) % was evidently higher than those in untreated group(45.64±1.26)% and control siRNA group(46.23±1.39)%,and there was a significant difference(P<0.05).Additionally,percentage of cell number in S phase in HDAC6 siRNA group(25.93±0.73)% was significantly lower than those in untreated group(33.13±1.02)% and control siRNA group(32.98±0.91)%(P<0.05).CONCLUSIONS: HDAC6 siRNA can effectively downregulate the expressions of HDAC6 mRNA and protein in ESCC cell line EC9706 cells.Downregulation of HDAC6 expression can obviously inhibit cell proliferation of ESCC cell line EC9706 cells,arrests cell cycle at G0/G1 phase.更多还原