To Investigate The Mechanism Of "?-hCG-ERK1/2-MMP-2" Signal Pathway In Ovarian Cancer Invasion And Metastasis

Background/Object: Ovarian cancer carries a high mortality due to its properties of invasion and metastatic potential,however,underlying molecular events remain elusive.Recently,it was shown that?-h CG,which has a high expression level in a variety of malignant tumors,plays an important role in the process of tumor invasion and metastasis.The synergistic effect of extracellular signal-regulated kinase(ERK1/2)and its downstream matrix metalloproteinases(MMPs)is the key link of tumor cell migration and invasion.This study aims to investigate the role and mechanism of "?-h CG-ERK1/2-MMP-2" signaling pathway in ovarian cancer invasion and metastasis.Methods: The lentiviral LV-?-hCG and si RNA-?-hCG were transfected into human epithelial ovarian cancer(EOC)cells,while LV-control and si RNA-NC were used as negative control.q RT-PCR and western blot were used to compare the expression of ?-h CG in each group and verify the transfection efficiency.The wound healing assay,transwell migration and invasion assays were performed to investigate the effects of ?-h CG on migration and invasion ability in human EOC cells.To explore the role and mechanism of ?-h CG in ovarian cancer metastasis,q RT-PCR and western blot were used to detect the expression of ?-h CG,ERK1/2,p-ERK1/2,MMP-2.si RNA-ERK1/2 and ERK1/2 phosphorylation inhibitor SCH772984 were transfected into the over-expressing ?-h CG human EOC cells,si RNA-NC and DMSO were used as negative control.Transwell migration and invasion assays were used to investigate the effect of over-expressed ?-h CG with “rescue” of ERK1/2 on migration and invasion ability of human EOC cells in vitro.q RT-PCR and western blot were used to detect the expression of ERK1/2,p-ERK1/2,MMP-2.Statistical analyses were performed using Graph Pad Prism 6,t-test was utilized to analyze,all data are presented as "mean ± standard deviation",the difference was statistically significant(P <0.05).All experiments were repeated 3 times.Results: In this study,overexpression and knockdown of ?-hCG ovarian cancer cell model were constructed successfully.The migration and invasion ability of EOC cells,which was detected by wound healing assay,transwell migration and invasion assays,were significantly facilitated by up-regulation of ?-h CG in vitro,while the silencing ?-h CG led to the opposite effect,the difference was statistically significant(P <0.05).Moreover,the expression of ERK1/2,p-ERK1/2 and MMP-2 in EOC cells were increased or decreased accordingly mediated by up-or down-regulated ?-h CG expression,and the difference was statistically significant(P <0.01).Furthermore,the results of transwell migration and invasion assays showed that migration and invasion ability was decreased in EOC cells when overexpression of ?-h CG with “rescue” of ERK1/2 in vitro,and the expression of p-ERK1/2 and MMP-2 in EOC cells were decreased by down-regulated ERK1/2 expression while up-regulated ?-h CG detected by q RT-PCR and western blot,the difference was statistically significant(P<0.001).Conclusion: ?-hCG plays an important role in regulating the invasion and metastasis via the "ERK12-MMP-2" signaling pathway in ovarian cancer,which may be a potential target for treatment of ovarian cancer.