Abstract:
Abstract:Objective To explore the effects of AKT1 gene on the invasion and migration of epithelial ovarian cancer cells SKOV3 and its mechanism. Methods Recombinant plasmid carrying the full-length AKT1 cDNA and AKT1 shRNA plasmid were constructed. Plasmids were transfected into SKOV3 cells to dual-directionally regulate AKT1 expression. RT-PCR and western blot were used to detect the transfection efficiency. Wound healing assay was used to analyze cell migration. Transwell-Matrigel assay was used to analyze cell invasion. RT-PCR was used to explore the expression of CXCR4, VEGF, MMP-2, MMP-9 and uPA which were related with cell invasion and migration at mRNA level. Results A plasmid (pEF-1α-AKT1) containing full length of AKT1 cDNA and a specifi c AKT1-targeted shRNA expression plasmid were constructed successfully, which could effectively regulate the activation of p-AKT after SKOV3 cells transfected. Compared with untranfected cells or non-target shRNA transfected cells, up-regulation AKT1 promoted cell migration and invasion, and increased the expression of CXCR4, VEGF, MMP-2, and uPA at mRNA level(P<0.05). On the other hand, down-regulation AKT1 inhibited cell migration and invasion, and decreased the expression of CXCR4, VEGF, MMP-2, and uPA at mRNA level(P<0.05). Conclusion AKT1 might impact cell migration and invasion by regulating the expression of CXCR4, VEGF, MMP-2, and uPA at mRNA level.