miR-142-3p在MCF-7细胞中靶向调节AKT pathway活性机理

为探究miR-142-3p在MCF-7细胞中作用机理,采用RNA免疫共沉淀技术和双荧光素酶报告基因技术筛选及验证miR-142-3p作用靶基因;蛋白质免疫印迹技术验证靶基因及其介导的PI3K-AKT-mTOR信号通路蛋白表达量及通路活性;应用实时荧光定量PCR验证靶基因PTEN对miR-142-3p调节关系。结果表明,miR-142-3p靶向调节AKT和PTEN表达;miR-142-3p过表达组中PI3K-AKT-mTOR通路活性显著降低;miR-142-3p抑制组中PI3K-AKT-mTOR通路活性显著上升;抑制PTEN表达显著提高miR-142-3p表达量。因此miR-142-3p靶向调节AKT和PTEN表达继而抑制PI3K-AKT-mTOR信号通路活性,PTEN与miR-142-3p存在调节关系。

Regulating mechanism of miR-142-3p inhibit PI3K-AKT-mTOR signaling pathway in MCF-7 cell line

To analyze the mechanism of miR-142-3p in MCF-7 cell line. In this research, RNA coimmunoprecipitation and dual luciferase reporter gene technology were used to screen and validate target genes of miR-142-3p. Western blot was used to verify the expression of proteins in PI3K-AKTmTOR pathway after miR-142-3p was overexpressed or inhibited in breast cancer cell MCF-7. Realtime RT-PCR verified the regulation relationship of target gene PTEN of miR-142-3p. The dates showed that miR-142-3p regulated the expression of AKT and PTEN; the activity of PI3K-AKT-mTOR pathway was significantly decreased in the miR-142-3p over expression group; the activity of PI3K-AKT-mTOR pathway was significantly increased in the inhibition group; Inhibition of PTEN expression significantly increased the expression of miR-142-3p. Conclusion:miR-142-3 p inhibit the activity of PI3K-AKT-mTOR signalling pathway by regulating the expression of AKT and PTEN, and PTEN had a regulatory relationship with miR-142-3 p.