本文選題：宮頸癌 + miR-�。� 參考：《中華腫瘤防治雜志》2015年20期

【摘要】：目的 microRNA-218(miR-218)在多種腫瘤中低表達(dá),是潛在的腫瘤抑制因子,本研究探討miR-218過(guò)表達(dá)對(duì)宮頸癌細(xì)胞增殖和遷移的影響。方法構(gòu)建miR-218慢病毒表達(dá)載體并感染宮頸癌SiHa、HeLa和C-33A細(xì)胞,篩選出穩(wěn)定表達(dá)miR-218的細(xì)胞株;通過(guò)熒光定量PCR(qRT-PCR)檢測(cè)轉(zhuǎn)染后3株細(xì)胞miR-218、miR-9、miR-21和miR-31的表達(dá);克隆形成實(shí)驗(yàn)和MTT實(shí)驗(yàn)檢測(cè)細(xì)胞的增殖情況;劃痕實(shí)驗(yàn)及Transwell細(xì)胞遷移實(shí)驗(yàn)檢測(cè)細(xì)胞的遷移能力;qRT-PCR檢測(cè)預(yù)測(cè)靶基因LAMB3和Robo1mRNA的表達(dá)。結(jié)果成功構(gòu)建miR-218慢病毒表達(dá)載體。與對(duì)照組相比,感染miR-218慢病毒的3株宮頸癌細(xì)胞SiHa、HeLa和C-33A中miR-218的表達(dá)量明顯升高,而miR-9、miR-21和miR-31的表達(dá)沒(méi)有明顯變化;實(shí)驗(yàn)組(過(guò)表達(dá)miR-218)與對(duì)照組相比可降低SiHa(t=10.73,P=0.008 6)和C-33A(t=4.992,P=0.037 9)細(xì)胞的克隆形成率;MTT結(jié)果表明,實(shí)驗(yàn)組與對(duì)照組相比,miR-218對(duì)SiHa細(xì)胞增殖抑制作用第3天最明顯(t=50.46,P=0.000 4),對(duì)C-33A細(xì)胞增殖抑制作用第4天最明顯(t=16.13,P=0.003 8),而對(duì)HeLa的克隆形成能力和增殖能力均無(wú)影響;遷移實(shí)驗(yàn)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)的劃痕愈合率,并且可以減少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)的穿膜細(xì)胞數(shù)。此外,過(guò)表達(dá)miR-218可明顯抑制宮頸癌細(xì)胞中LAMB3和Robo1基因mRNA的表達(dá)。結(jié)論 miR-218可抑制宮頸癌細(xì)胞的增殖和遷移,可能參與了宮頸癌的發(fā)生和發(fā)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.
【作者單位】： 武漢大學(xué)人民醫(yī)院婦科;廣西壯族自治區(qū)人民醫(yī)院婦科;廣西壯族自治區(qū)人民醫(yī)院科研實(shí)驗(yàn)中心;
【基金】：國(guó)家自然科學(xué)基金(81060219) 廣西自然科學(xué)基金(2014GXNSFAA118266)
【分類(lèi)號(hào)】：R737.33

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本文編號(hào)：2092008