微小RNA-148a抑制乳腺癌细胞MDA-MB-231迁移的作用机制

目的:研究微小RNA-148a(miR-148a)在乳腺癌细胞株MDA-MB-231中的表达,探讨上调miR-148a的表达对其迁移的影响及机制.方法:采用实时荧光定量PCR(qRT-PCR)检测正常乳腺上皮细胞MCF-10A和乳腺癌细胞株MDA-MB-231中miR-148a的表达水平.应用脂质体法将miR-148a mimic及阴性对照分别转染MDA-MB-231细胞,qRT-PCR检测转染效率;细胞划痕实验检测转染前后MDA-MB-231细胞迁移能力的变化;Western blot方法检测上皮间质转化(EMT)相关蛋白Slug、Snail和E-cadher-in表达水平的变化.结果:与MCF-10A细胞相比,MDA-MB-231细胞中miR-148a的表达水平明显降低(P<0.05);与阴性对照组相比,miR-148a转染组中miR-148a的表达水平显著升高(P<0.05);过表达miR-148a后,MDA-MB-231细胞迁移能力明显下降,Slug和Snail蛋白表达明显降低,E-cadherin蛋白表达明显增加.结论:miR-148a可通过调控Slug、Snail及E-cadherin蛋白的表达抑制EMT,进而抑制乳腺癌细胞的迁移能力.

The mechanisms of miR - 148a inhibiting migration of breast cancer MDA - MB - 231 cells

Objective:To study the expression of miR - 148a in MDA - MB - 231 breast cancer cells,and to ex-plore the effect and mechanism of miR - 148a on the breast cancer cells migration. Methods:Quantitative real - time PCR (qRT - PCR)was used to detect the expression of miR - 148a in MCF - 10A cells and human breast cancer cell line MDA - MB - 231. miR - 148a mimic and negative control were transfected into MDA - MB - 231 cells. The efficiency of transfection was measured by qRT - PCR. Cell scratch assay was used to observe the migration ability changes. Western blot was performed to detect the expression levels of epithelial - mesenchymal transition (EMT)re-lated proteins Slug,Snail and E - cadherin. Results:The expression level of miR - 148a in MDA - MB - 231 cells de-creased significantly (P < 0. 05). miR - 148a mimic obviously increased the expression of miR - 148a,inhibited the cell migration,decreased Slug and Snail proteins expression,increased E - cadherin protein expression in MDA - MB- 231 cells (P < 0. 05). Conclusion:miR - 148a may inhibit EMT and migration by regulating the expression of Slug,Snail and E - cadherin of breast cancer cells.