敲低HSP60 对结肠癌SW480 细胞增殖能力的影响及其机制探究

目的:研究热休克蛋白60(HSP60)敲低对结肠癌SW480细胞增殖的影响,并进一步探究其作用机制。方法:通过含HSP60sh RNA载体的慢病毒感染加上流式细胞仪无菌分选的方法构建结肠SW480 HSP60基因稳定RNA干扰(RNAi)单克隆细胞系,利用Western blot和q-PCR验证结肠癌细胞中HSP60的敲低效率;使用CCK-8试剂检测结肠癌细胞增殖能力,并用流式细胞仪检测其HSP60敲低对细胞周期的影响。结果:Western blot和q-PCR结果验证了HSP60在结肠癌细胞中的敲低效率,与对照组细胞相比,实验组细胞HSP60的m RNA水平和蛋白水平均降低了60%以上。CCK-8实验结果表明,敲低HSP60后SW480细胞的增殖能力下降了约70%;流式细胞周期实验显示敲低HSP60后SW480细胞中G0/G1期、S期、G2/M期的分布比例变化不大。结论:敲低HSP60能够显著抑制SW480细胞的增殖能力,而SW480细胞周期并没有发生明显变化,推测HSP60的敲低引起的线粒体损伤导致细胞生长速度变慢。

The Effects of HSP60 Knockdown on the Proliferation of Colon Cancer SW480 and its Mechanism

Objective:To investigate the effects of HSP60 knockdown on the proliferation of colon cancer SW480 cells.Methods:Lentivirus containing the HSP60-directed shRNAs was used to construct a stable monoclonal HSP60 knockdown cell line in SW480. The HSP60 knockdown efficiency was determined by western blotting and the theCCK-8 assay was used to analyze cell proliferation rates, while flow cytometry was used to determine the cell cycle in HSP60-knocked down cells.Results:We established a stable cell line in which HSP60 expression was silenced in SW480 cells as confirmed by western blotting and q-PCR. CCK-8 assay showed that HSP60 knockdown significantly inhibited the cell proliferation, whereas it had little effect on the cell cycle progression.Conclusion:HSP60 knockdown inhibits the cell proliferation in SW480 cells. so we speculate that the reduce of SW480 growing is due to mitochondrial damage.