miR-149-5P调控β-catenin/MMP2/E-cadherin介导胃癌BGC-823和MKN28的细胞迁移

目的 观察miR-149-5P对胃癌细胞BGC-823和MKN28迁移的影响。方法 采用qRT-PCR方法检测miR-149-5P在胃粘膜上皮细胞(GES-1)与BGC-823和MKN28细胞中的表达。分别转染miR-149-5P mimics、NC、inhibitor和inhibitor-NC,采用qRT-PCR方法验证转染效率,采用细胞划痕实验和Transwell实验检测过表达或抑制miR-149-5P后胃癌细胞BGC-823和MKN28迁移能力。采用Western Blot检测转染miR-149-5P mimics或inhibitor后BGC-823和MKN28细胞中β-catenin、MMP2、E-cadherin蛋白表达水平。结果 qRT-PCR实验结果显示:与GES-1相比,miR-149-5P在BGC-823和MKN28细胞中表达显著降低;转染mimics后,BGC-823和MKN28细胞中miR-149-5P表达水平显著升高,而转染inhibitor后,BGC-823和MKN28细胞中miR-149-5P表达水平下降。细胞划痕实验和Transwell实验结果显示:转染mimics后可以抑制BGC-823和MKN28细胞迁移,而转染inhibitor后,可以促进BGC-823细胞迁移,MKN28细胞并无明显变化。Western Blot实验结果显示:在BGC-823和MKN28细胞中过表达miR-149-5P可上调E-cadherin的蛋白表达,抑制β-catenin和MMP2蛋白表达,而抑制miR-149-5P的表达可下调E-cadherin的蛋白表达,促进BGC-823细胞中β-catenin和MMP2蛋白表达,但转染inhibitor后MKN28细胞中E-cadherin、β-catenin和MMP2蛋白表达水平并无明显变化。结论 miR-149-5P可以抑制胃癌细胞BGC-823和MKN28迁移。

miR-149-5p regulates β-catenin/MMP2/E-cadherin mediated cell migration of BGC-823 and MKN28 in gastric cancer

Objective To investigate the effect of miR-149-5P on the migration of gastric cancer cells BGC823 and MKN28.Methods The expression of miR-149-5P in GES-1 cells and BGC-823 cells and MKN28 cells was detected by qRT-PCR.the miR-149-5P mimics,NC,inhibitor and inhibitor-NC were transfected into BGC-823 cells and MKN28 cells separately,and the efficiency of transfection was also determined by qRT-PCR.The migration ability of BGC-823 and MKN28 in gastric cancer cells after over expression or inhibition of miR-149 was measured by cell Transwell and scratch assay.Western blot was used to detect the expression of β-catenin,MMP2,E-cadherin BGC-823 cells and MKN28 cells transfected with miR-149-5P mimics or inhibitor.Results Compared with GES-1,the expression of miR-149-5P in BGC-823 and MKN28 cells was decreased.After mimics transfected,the expression level of miR-149-5P in BGC-823 and MKN28 cells was higher than NC group.However,compared with inhibitor-NC,the inhibitor group was decreased.The results of cell scratch test and Transwell test showed that the migration of BGC-823 and MKN28 cells could be blocked by transfection of mimics,after transfection with inhibitor,the migration of BGC-823 cells was promoted,but MKN28 cells did not change significantly.Western blot showed that overexpression of miR-149-5p in BGC-823 and MKN28 cells could up regulate the expression of E-cadherin,inhibit the expression of β-catenin and MMP2,while inhibition of miR-149-5p could down regulate the protein expression of E-cadherin and promote the protein expression of β-catenin and MMP2 in BGC-823 cells,However,the expression levels of E-cadherin,β-catenin and MMP2 in MKN28 cells did not change significantly after transfection.Conclusion miR-149-5P can inhibit the migration of BGC-823 and MKN28 cells.