miR-138靶向调控FAK抑制三阴型乳腺癌MDA-MB-231细胞侵袭转移的机制

miR-138 suppresses migration and invasion of triple-negative breast cancer MDA-MB-231 cells by targeting FAK

目的探讨miR-138对三阴型乳腺癌(triple-negative breast cancer,TNBC)MDA-MB-231细胞侵袭、转移的影响及分子机制。方法将MDA-MB-231细胞转染miR-138 mimics及其阴性对照,Transwell实验检测细胞迁移和侵袭能力改变。生物信息学预测miR-138的潜在靶基因,采用双荧光素酶报告实验和Western blot法分析miR-138与FAK的靶向调控关系。回复实验使用Transwell及Western blot实验验证miR-138通过FAK抑制MDA-MB-231细胞侵袭、转移。结果Transwell实验结果表明:过表达miR-138后,MDA-MB-231细胞的迁移和侵袭能力均显著降低(P<0.05)。生物信息学预测FAK为miR-138的潜在靶基因,双荧光素酶报告实验结果表明:FAK 3′UTR区域存在miR-138的互补结合位点。Western blot结果发现过表达miR-138后FAK蛋白表达降低。回复实验Transwell及Western blot结果表明:上调FAK可以部分恢复miR-138过表达所抑制的细胞迁移和侵袭能力(P<0.05);使用FAK siRNA敲低FAK可以部分恢复miR-138抑制所增强的细胞迁移与侵袭能力(P<0.05)。结论miR-138靶向调控FAK抑制TNBC细胞的侵袭、转移。

Purpose To investigate the mechanism of miR-138 on metastasis of triple-negative breast cancer(TNBC)MDA-MB-231 cells.Methods Transwell assay was used to detect cell migration and invasion ability after MDA-MB-231 cells were transfected with miR-138 mimics and its negative control.Bioinformatics was used to predict potential target genes of miR-138.Luciferase assay and Western blot were used to verify the targeting relationship between miR-138 and FAK.Rescue assay was used to verified whether miR-138 inhibited metastasis of TNBC cells through FAK by Transwell and Western blot.Results Transwell assay results showed that overexpression of miR-138 inhibited the migration and invasion of MDA-MB-231 cells(P<0.05).FAK was predicted as the target of miR-138 by bioinformatics.In addition,the target relationship was verified between miR-138 and FAK by luciferase reporter plasmids assay.Western blot result showed that the expression level of FAK was significantly inhibited after the overexpression of miR-138.Furthermore,Transwell and Western blot results of function-rescue assay showed that the restoration of the expression of FAK could partially restore the cell migration and invasion ability inhibited by miR-138 overexpression.Knockdown of FAK by siRNA could partially restore the cell migration and invasion ability enhanced by miR-138 inhibition(P<0.05).Conclusion MiR-138 negatively regulates the expression of FAK to inhibit the TNBC metastasis.