MicroRNA-155 Induced Invasion And Migration Of Human Trophoblast Cells Via CXCR4/PI3K/AKT Signaling Pathway

ObjectiveTo 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.Materials and Methods 1 MaterialsIn this study,we selected human choriocarcinoma cell line JEG-3 cell line,its biological traits close to the original isolated trophoblast cells,and the cell line invasiveness and early pregnancy trophoblast no significant difference.2 MethodsChemically 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.3 Statistics analysisAll the data in this study were analyzed by SPSS21.0 software,and the results were expressed asx ± s.Comparison between the two groups using two independent samples t test,multi-group comparison using one-way analysis of variance.With ? = 0.05 for the test level.Results 1 The relative expression of CXCR4 mRNA of JEG-3 after the transfection of miR-155The result of RT-PCR showed that the expression of CXCR4 mRNA in JEG-3 cells was lower in miR-155 mimics group(0.589±0.096)compared to that in the blank control group(1.503±0.090)and negative control group(1.146±0.153).The difference was statistically significant(F=36.59,P=0.035);compared to the other two group,the expression of CXCR4 mRNA in JEG-3 cells,was higher in miR-155 inhibitor group(1.739±0.083).The difference was statistically significant(F=28.34,P=0.027).2 The relative expression level of CXCR4 and p-AKT protein of JEG-3 after the transfection of miR-155The result of Western blot showed that the expression of CXCR4 and p-AKT protein of JEG-3 cells was down-regulated after transfected miR-155 mimics.The difference was statistically significant(F=126.67,P=0.000;F=114.83,P=0.000).And the expression of CXCR4 and p-AKT protein of JEG-3 cells was up-regulated after transfected miR-155 inhibitor.The difference was statistically significant(F=16.85,P=0.002;F=14.72,P=0.003).3 Effect on the ability of invasion of JEG-3 cells after the transfection of miR-155The result of transwell invasion assay showed that the number of transmembrane cells was less in miR-155 mimics group(22.89±9.42)than that in the blank control group(63.46±2.37)and negative control group(49.29±5.81).The difference was statistically significant(F=127.37,P=0.000);compared to the other two group,the number of transmembrane cells was more in miR-155 inhibitor group(81.50±11.25).The difference was statistically significant(F=2.94,P=0.027).4 Changes of migration distance of JEG-3 cells after the transfection of miR-155The result of wound healing assay showed that the relative migration distance was decreased in miR-155 mimics group(0.159±0.058)compared to the blank control group(1.080 ± 0.045)and negative control group(0.823 ± 0.201).The difference was statistically significant(F=29.600,P=0.017);compared to the other two group,the relative migration distance was decreased in miR-155 inhibitor group(1.640 ± 0.078).The difference was statistically significant(F=16.103,P=0.024).ConclusionsmiR-155 may inhibits the invasion and migration of trophoblast cells by regulating CXCR4/PI3K/AKT pathway contributing to the development of preeclampsia.