microRNA-218对宫颈癌细胞增殖和迁移的影响

发布时间：2018-07-03 02:36

本文选题：宫颈癌 + miR-　；参考：《中华肿瘤防治杂志》2015年20期

【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。
[Abstract]:Objective to investigate the effect of miR-218 overexpression on the proliferation and migration of cervical cancer cells. Methods the expression vector of miR-218 lentivirus was constructed and infected with SiHa-HeLa and C-33A cells, and the stable expression of miR-218 was screened, and the expression of miR-218- miR-9miR-21 and miR-31 was detected by fluorescence quantitative PCR (qRT-PCR). Cell proliferation was detected by clone formation assay and MTT assay, and cell migration ability was detected by scratch assay and Transwell cell migration assay. The expression of LAMB3 and Robo1 mRNA was detected by qRT-PCR. Results the expression vector of miR-218 lentivirus was successfully constructed. Compared with the control group, the expression of miR-218 in the three cervical cancer cells infected with miR-218 lentivirus was significantly higher than that in the control group, but the expression of miR-21 and miR-31 in the three cervical cancer cells infected with miR-218 was not significantly changed. Compared with the control group, the overexpression of miR-218 in the experimental group decreased the colony formation rate of SIHA (tH10.73) P0. 0086 and C-33A (t0. 992P0. 037 9) cells. Compared with the control group, the inhibitory effect of miR-218 on SIHA cell proliferation was the most obvious on the 3rd day (tnr 50.46U Pu 0.0004), and on the C-33A cell line on the 4th day (tr 16.13PnP 0.0038), but it had no effect on the clone forming ability and proliferation ability of HeLa. Migration experiments showed that miR-218 could inhibit the rate of scratch healing in SIHA (tHH 12.90 Pn0. 0060) HeLa (tnl10.70P0. 0086) and C-33A (T5. 099P0. 036 4), and could reduce the number of perforated cells of SIHA (tl. 13.75P0. 0052) HeLa (tl. 8. 643Pn0. 0131) and C-33A (t15. 10TnP0. 0044). In addition, overexpression of miR-218 could significantly inhibit the expression of LAMB3 and Robo1 mRNA in cervical cancer cells. Conclusion miR-218 can inhibit the proliferation and migration of cervical cancer cells and may be involved in the occurrence and development of cervical cancer.
【作者单位】：武汉大学人民医院妇科;广西壮族自治区人民医院妇科;广西壮族自治区人民医院科研实验中心;
【基金】：国家自然科学基金(81060219) 广西自然科学基金(2014GXNSFAA118266)
【分类号】：R737.33

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