MicroRNA-155通过调节CXCR4/PI3K/AKT途径影响滋养细胞的侵袭与迁移

MicroRNA-155 induced invasion and migration of human trophoblast cells via CXCR4/PI3K/AKT signaling pathway

目的:以人绒毛膜滋养层细胞系JEG-3细胞为研究对象,结合该细胞侵袭和迁移能力的变化情况,研究转染miR-155 mimics和miR-155 inhibitor之后CXCR4的表达变化及其对下游PI3K/AKT信号通路的影响作用,从而探讨miR-155参与子痫前期发生发展的分子机制。方法:设计miR-155 mimics和miR-155 inhibitor,对JEG-3进行转染,通过Transwell侵袭实验、划痕实验,观察转染后细胞的侵袭和迁移能力的变化;利用Real-time PCR检测CXCR4 mRNA的表达;利用Western blot检测CXCR4及下游p-AKT蛋白的表达水平。结果:Real-time PCR结果显示,miR-155 mimics转染组CXCR4 mRNA相对表达量(0.589±0.096)明显低于空白对照组(1.503±0.090)和阴性对照组(1.146±0.153),差异有统计学意义(P<0.05);miR-155 inhibitor转染组CXCR4 mRNA相对表达量(1.739±0.083)与两组对照组相比差异也有统计学意义(P<0.05)。Western blot结果显示miR-155 mimics转染组CXCR4蛋白和下游p-AKT蛋白表达水平均明显降低,而miR-155 inhibitor转染组CXCR4和p-AKT蛋白水平则升高,与空白对照组和阴性对照组相比差异均有统计学意义(P<0.05)。Transwell侵袭实验结果显示,与空白对照组(63.46±2.37)和阴性对照组(49.29±5.81)侵袭细胞数相比,miR-155 mimics转染组侵袭细胞数(22.89±9.42)明显减少,差异有统计学意义(P<0.05);同时,miR-155 inhibitor转染组侵袭细胞数(81.50±11.25)明显升高,差异有统计学意义(P<0.05)。划痕实验结果显示,转染miR-155 mimics后,JEG-3细胞相对迁移距离(0.159±0.058)低于空白对照组(1.080±0.045)和阴性对照组(0.823±0.201),差异有统计学意义(P<0.05);而miR-155 inhibitor转染组,JEG-3细胞相对迁移距离(1.640±0.078)明显增加,差异有统计学意义(P<0.05)。结论:miR-155可能通过抑制CXCR4的表达进而抑制其下游PI3K/AKT信号通路的活化,从而影响滋养细胞的侵袭及迁移能力,最终导致子痫前期的发生发展。

Objective： To investigate the effect on the CXCR4 /PI3K/AKT pathway after the transfection of miR-155 mimics and miR-155 inhibitor combined with the research on the ability of invasion and migration of human chorionic JEG-3 trophoblast cells. Methods： Chemically synthesized miR-155 mimics and miR-155 inhibitor were transfected into JEG-3 cells. The effect on the ability of invasion and migration were analyzed by Transwell migration assay and Wound healing assay. The expression of CXCR4 mRNA was detected by Real-time PCR. The expression of CXCR4 and p-AKT protein were detected by Western blot. Results： Transfection with miR-155 mimics significantly down-regulated the expression of CXCR4 as compared with the control group（ P〈0. 05）; JEG-3 cells transfected miR-155 mimics had lower levels of migration and invasion capacity than cells in the control group（ P〈0. 05）. However,transfection with miR-155 inhibitor significantly up-regulated the expression of CXCR4 as compared with the control group（ P〈0. 05）;JEG-3 cells transfected miR-155 inhibitor had higher levels of migration and invasion capacity than cells in the control group（ P〈0. 05）. Addition,the expression of p-AKT protein of JEG-3 cells was down-regulated after transfected miR-155 mimics,and the expression of p-AKT protein of JEG-3 cells was up-regulated after transfected miR-155 inhibitor. Conclusion： miR-155 may inhibits the invasion and migration of trophoblast cells by regulating CXCR4 / PI3 K / AKT pathway contributing to the development of preeclampsia.