microRNA-218对宫颈癌细胞增殖和迁移的影响  

Effect of microRNA-218 on the proliferation and migration of cervical cancer cells

作　　者：刘珍[1,2] 张蔚[1] 胡晓霞[2] 蓝娇[3] 肖瑞平[3] 陈璐璐[2] 王琳琳[2] 

机构地区：[1]武汉大学人民医院妇科,湖北武汉430000 [2]广西壮族自治区人民医院妇科,广西南宁530000 [3]广西壮族自治区人民医院科研实验中心,广西南宁530000

出　　处：《中华肿瘤防治杂志》2015年第20期1602-1608,共7页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(81060219);广西自然科学基金(2014GXNSFAA118266)

摘　　要：目的 microRNA-218(miR-218)在多种肿瘤中低表达,是潜在的肿瘤抑制因子,本研究探讨miR-218过表达对宫颈癌细胞增殖和迁移的影响。方法构建miR-218慢病毒表达载体并感染宫颈癌SiHa、HeLa和C-33A细胞,筛选出稳定表达miR-218的细胞株;通过荧光定量PCR(qRT-PCR)检测转染后3株细胞miR-218、miR-9、miR-21和miR-31的表达;克隆形成实验和MTT实验检测细胞的增殖情况;划痕实验及Transwell细胞迁移实验检测细胞的迁移能力;qRT-PCR检测预测靶基因LAMB3和Robo1mRNA的表达。结果成功构建miR-218慢病毒表达载体。与对照组相比,感染miR-218慢病毒的3株宫颈癌细胞SiHa、HeLa和C-33A中miR-218的表达量明显升高,而miR-9、miR-21和miR-31的表达没有明显变化;实验组(过表达miR-218)与对照组相比可降低SiHa(t=10.73,P=0.008 6)和C-33A(t=4.992,P=0.037 9)细胞的克隆形成率;MTT结果表明,实验组与对照组相比,miR-218对SiHa细胞增殖抑制作用第3天最明显(t=50.46,P=0.000 4),对C-33A细胞增殖抑制作用第4天最明显(t=16.13,P=0.003 8),而对HeLa的克隆形成能力和增殖能力均无影响;迁移实验表明,miR-218可抑制SiHa(t=12.90,P=0.006 0)、HeLa(t=10.70,P=0.008 6)和C-33A(t=5.099,P=0.036 4)的划痕愈合率,并且可以减少SiHa(t=13.75,P=0.005 2)、HeLa(t=8.643,P=0.013 1)和C-33A(t=15.10,P=0.004 4)的穿膜细胞数。此外,过表达miR-218可明显抑制宫颈癌细胞中LAMB3和Robo1基因mRNA的表达。结论 miR-218可抑制宫颈癌细胞的增殖和迁移,可能参与了宫颈癌的发生和发展。OBJECTIVE MicroRNA 218 （miR-218） is a potential tumor suppressor and low expression in a variety of tumors, this study is to investigate the effect of overexpression of miR-218 on the proliferation and migration of cervical cancer cells. METHODS The miR-218 lentiviral vector was constructed and infected into cervical cancer SiHa, HeLa and C 33a cells. The cell lines with miR 218 expression were screened. The expressions of miR-218, miR-9, miR 21 and miR 31 were detected in the three kinds of cells after infecting by fluorescence quantitative PCR （qRT-PCR）. The cell proliferation was detected by clone formation assay and MTT assay. The cell migration ability was detected by scratch test and Transwell cell migration assay. The expressions of predicted target genes LAMB3 and Robol mRNA were detected by qRT-PCR. RESULTS The miR-218 lentiviral expression vector was successfully constructed. Compared with the control group, the expression of miR 218 was increased significantly in the three kinds of cervical carcinoma cells SiHa, HeLa and C-33A which were infected by the miR-218 lentiviral vector. There was no significant change in the expressions of miR-9, miR-21 and miR-31. Compared with the control group, the clone formation rates of SiHa （t= 10. 73, P= 0.008 6） and C33A（t=4. 992, P=0. 037 9） cells were decreased in miR-218 overexpression group. MTT results showed that, compared with the control group, the anti-proliferative effect of miR 218 on SiHa was the most significant on the third day （t=50.46, P=0. 000 4）, the anti-proliferative effect of miR-218 on C-33A was the most significant on the fourth day （t= 16.13, P=0. 003 8）, but miR-218 had no effect on clone formation ability and proliferation ability of HeLa. The migration experiment showed that miR-218 could inhibit the scratch healing rates of SiHa （t= 12.90, P= 0. 006 0）, HeLa （t=10.70, P=0. 008 6） and C-33A （t=5. 099, P=0. 036 4） and reduce the transmembrane cells of Si- Ha （t=13.75, P=0.005 2）, HeLa （t=8.643, P=0.0

关 键 词：宫颈癌 MIR-218 增殖 迁移 

分 类 号：R737.33[医药卫生—肿瘤]