摘要
目的探讨微小RNA(miR)-138通过调控上皮间质转化(EMT)影响乳腺癌细胞侵袭和迁移的分子机制。方法2017年7月至2018年6月采用逆转录-聚合酶链反应(RT-PCR)检测正常乳腺上皮细胞MCF-10A和乳腺癌细胞MCF-7、MDA-MB-231转染miR阴性对照模拟物(miR-NC)和miR-138模拟物后miR-138表达量;MTT法检测乳腺癌细胞活性;细胞划痕实验和Transwell实验分别检测细胞迁移距离和侵袭率。RT-PCR检测转染miR-138模拟物后EMT关键分子Vimentin、N-cadherin和E-cadherinmRNA表达量。结果MCF-10A中miR-138表达水平明显高于MCF-7和MDA-MB-231(1.006±0.009比0.324±0.027和0.512±0.068),差异有统计学意义(P<0.05);MCF-7和MDA-MB-231miR-138表达水平比较差异无统计学意义(P>0.05)。MCF-7和MDA-MB-231转染miR-138模拟物后第3和4天乳腺癌细胞活性明显低于转染miR-NC(MCF-7:0.514±0.052比0.593±0.061和0.643±0.074比0.784±0.081;MDA-MB-231:0.552±0.043比0.614±0.063和0.673±0.074比0.792±0.077),差异有统计学意义(P<0.05)。MCF-7和MDA-MB-231转染miR-138模拟物后乳腺癌细胞迁移距离和侵袭率明显低于转染miR-NC(MCF-7:0.572±0.051比1.003±0.012和0.624±0.043比1.002±0.007,MDA-MB-231:0.472±0.051比1.003±0.095和0.573±0.044比1.004±0.091),差异有统计学意义(P<0.05)。MCF-7和MDA-MB-231转染miR-138模拟物后Vimentin和N-cadherinmRNA表达量明显低于转染miR-NC,而E-cadherinmRNA表达明显升高,差异有统计学意义(P<0.05)。结论miR-138在乳腺癌细胞中表达量降低;miR-138可以抑制乳腺癌细胞增殖,并通过调控EMT过程抑制细胞侵袭和迁移能力。
ObjectiveTo investigate the molecule mechanism of microRNA (miR)-138 in inhibiting invasion and migration of breast cancer by regulating epithelial mesenchymal transformation (EMT).MethodsReverse transcription-polymerase chain reaction (RT-PCR) was used to detect expression of miR-138 after transfecting miR negative control simulacrum (miR-NC) and miR-138 simulacrum in human normal mammary epithelial cell (MCF-10A) and breast cancer cells (MCF-7 and MDA-MB-231) from July 2017 to June 2018. MTT method was used to detect the breast cancer cell activity. Cell scratch test and Transwell test were used to detect the breast cancer cell migration distance and invasion rate. RT-PCR was used to detect the expression of the EMT key molecules Vimentin, N-cadherin and E-cadherin after transfecting miR-138 simulacrum.ResultsThe expression level of miR-138 in MCF-10A was significantly higher than that in MCF-7 and MDA-MB-231 (1.006 ± 0.009 vs. 0.324 ± 0.027 and 0.512 ± 0.068), and there was statistical difference (P<0.05);there was no statistical difference in the expression level of miR-138 between MCF-7 and MDA-MB-231 (P>0.05). The breast cancer cell viabilities of MCF-7 and MDA-MB-231 at third and fourth day after transfecting miR-138 simulacrum were significantly lower than those of transfecting miR-NC (MCF-7: 0.514 ± 0.052 vs. 0.593 ± 0.061 and 0.643 ± 0.074 vs. 0.784 ± 0.081;MDA-MB-231: 0.552 ± 0.043 vs. 0.614 ± 0.063 and 0.673 ± 0.074 vs. 0.792 ± 0.077), and there were statistical differences (P<0.05). The breast cancer cell migration distances and invasion rates of MCF-7 and MDA-MB-231 after transfecting miR-138 simulacrum were significantly lower than those of transfecting miR-NC (MCF-7: 0.572 ± 0.051 vs. 1.003 ± 0.012 and 0.624 ± 0.043 vs. 1.002 ± 0.007, MDA-MB-231: 0.472 ± 0.051 vs. 1.003 ± 0.095 and 0.573 ± 0.044 vs. 1.004 ± 0.091), and there were statistical differences (P<0.05). The expressions of Vimentin and N-cadherin mRNA in MCF-7 and MDA-MB-231 after transfecting miR-138 simulacrum were significantly lower than those of transfecting miR-NC, but the expression of E-cadherin mRNA was significantly increased, and there were statistical differences (P<0.05).ConclusionsThe expressions of miR-138 in both breast cancer cells decreased. Overexpression of miR-138 in breast cancer cell can inhibit proliferation, migration and invasion via regulating EMT.
作者
沈庆林
彭敏
章必成
姚颐
徐唐鹏
褚玉新
宋启斌
Shen Qinglin;Peng Min;Zhang Bicheng;Yao Yi;Xu Tangpeng;Chu Yuxin;Song Qibin(Cancer Center,Renmin Hospital,Wuhan University,Wuhan 430060,China)
出处
《中国医师进修杂志》
2019年第1期42-46,共5页
Chinese Journal of Postgraduates of Medicine
基金
国家自然科学基金面上项目(81472748).
