RNA干扰抑制p62基因表达对宫颈癌HeLa细胞增殖的影响  

Effect of the silencing p62 gene expression by RNA interference on proliferation of cervical cancer HeLa cells

作　　者：陈推[1] 袁运水[1] 陈惠祯[2] 

机构地区：[1]湖北省通城县人民医院妇产科,437400 [2]武汉大学中南医院妇瘤科,湖北武汉430071

出　　处：《蚌埠医学院学报》2018年第1期13-15,共3页Journal of Bengbu Medical College

摘　　要：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。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.

关 键 词：宫颈肿瘤 短发夹RNA p62基因 宫颈癌HELA细胞 增殖 

分 类 号：R737.33[医药卫生—肿瘤]