Objective:To investigate the effects of downregulating the expression of p62 gene by shNA transfection on proliferation of cervical cancer HeLa cells. Methods:The lentiviral vector containing shRNA sequence of silencing p62 gene was constructed,and transfected into the cervical HeLa cells(shRNA group).The negative control group(shNC group) was set,and the non-lentiviral vector was set as blank control group.The expression levels of p62 mRNA,cell proliferation and cell colony formation rate in 3 groups were detected using qRT-PCR,CCK-8 assay and colony formation assay,respectively. Results:The lentiviral vector containing shRNA sequence of silencing p62 gene was successfully constructed.The expression of p62-mRNA in shRNA group and shNC group relative to the blank control group were (0.18 ± 0.08) and (1.01 ± 0.12),respectively.The interfere rate of shRNA was 81.00%.Compared with the negative control group and blank control group,the cell proliferation and colony formation rates in shRNA group were slow and little(P<0.05 and P<0.01),respectively.Conclusions:Silencing the expression of p62 gene can reduce the proliferation of cervical cancer HeLa cells.The p62 gene may play an important role in the proliferation of cervical cancer HeLa cells.%目的:研究shRNA降低p62基因表达对人宫颈腺癌HeLa细胞增殖的影响.方法:构建具有沉默p62表达的慢病毒载体短发夹RNA(shRNA)序列,采用Polybrene将其转染入宫颈腺癌HeLa细胞中(shRNA组),设立shNC作为阴性对照组,另设未加慢病毒转染载体的空白对照组.采用实时荧光定量qRT-PCR技术检测各组相对空白对照组的p62基因相对表达量;采用CCK-8实验检测3组细胞的增殖情况,采用集落克隆形成实验检测3组细胞的克隆形成率.结果:沉默p62基因表达的宫颈癌HeLa细胞成功构建;shRNA组p62mRNA相对空白对照组的相对表达量为(0.18 ± 0.08),而shNC组的p62mRNA相对空白对照组的相对表达量为(1.01 ± 0.12)(t=9.97,P<0.01);shRNA的干扰效率为81.00%;shRNA组较阴性对照组和空白对照组细胞增殖速度变慢(P<0.05),克隆形成率减少(P<0.01).结论:p62基因沉默后,宫颈癌HeLa细胞的增殖速度变慢.p62基因在宫颈癌HeLa细胞的增殖中起着重要的作用.
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