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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。
Objective: To investigate the effect of HDAC6 siRNA on HDAC6 expression in esophageal squamous carcinoma cell line EC9706 and the effect of HDAC6 expression on the proliferation and cell cycle of esophageal squamous cell carcinoma. Methods: The EC9706 cells were divided into three groups: untreated group, control siRNA group and HDAC6siRNA group. The latter two groups were respectively transfected with control siRNA and HDAC6siRNA. The effect of HDAC6siRNA on HDAC6 expression in esophageal squamous cell carcinoma EC9706 cells was detected by semi-quantitative reverse transcription-polymerase chain reaction, Western blotting and immunocytochemistry. The changes of cell proliferation were detected by CCK-8 kit. Flow cytometry was used to detect the changes of cell cycle in each group. Results: Compared with untreated group (0.951,0.924) and control group (0.947,0.905), the expression of HDAC6mRNA and HDAC6 mRNA in HDAC6siRNA group was 0.044, and the relative expression level of HDAC6 protein was 0.114 (T = 2.24 and 2.52 respectively, P <0.05). The cell proliferation was significantly inhibited and the ratio of G0 / G1 phase was (68.55 ± 2.01)%, which was significantly higher than that of untreated group (45.64 ± 1.26)% and control siRNA (46.23 ± 1.39)%, S phase (25.93 ± 0.73)% was significantly lower than that of untreated group (33.13 ± 1.02)% and control siRNA group (32.98 ± 0.91)%, the differences were statistically significant, P < 0.05. Conclusion: HDAC6siRNA can effectively down-regulate the expression of HDAC6mRNA and protein in esophageal squamous cell carcinoma EC9706 cells. The down-regulation of HDAC6 expression can significantly inhibit the proliferation of esophageal squamous carcinoma cell line EC9706 and induce cell cycle arrest in G0 / G1 phase.