The Study Of CSPG4 Promotes The Proliferation And The Migration Of Epithelial Ovarian Cancer Cells Through C-Met And ERK1/2

Objective:To study the effect of CSPG4 on the proliferation and migration of epithelial ovarian cancer cells,and to explore the activation of CSPG4 on its related oncogenic signaling pathways.Methods:1.Western Blot was used to determine the protein expression of CSPG4 in several invasive human ovarian cancer cell lines such as HEY,A2780,MA-148 and OVCAR-3 cell lines.CSPG4 stable interference and its empty vector cell lines(HEY sh RNA-CSPG4 and HEY sh RNA-control cells)were constructed using a cell transfection technique and in vitro functional assays were performed using these paired cell lines.Including: MTT assay and soft agar colony formation assay for cell proliferation,and scratch assay for cell migration.The importance of CSPG4 gene in the proliferation and migration of ovarian cancer cells was clarified.2.Determine the effect of CSPG4 expression on CSPG4-associated tumorigenic signaling pathway.The relevant pathway markers are: c-Met,Akt,ERK1 / 2.Interference with c-Met will be used to tentatively determine its potential importance as a downstream effector of CSPG4.Results:1.The protein expression of CSPG4 in HEY,OVCAR-3,A2780 and MA-148 was significantly higher than that of Sk OV3 and OVCAR-5.Western Blot results showed that HEY,OVCAR-3,A2780 and MA-148 were positive expression cell lines of CSPG4,and Sk OV3 and OVCAR-5 were negative expression cell lines of CSPG4.2.Immunofluorescence showed high expression of CSPG4 in ovarian cancer MA-148,HEY,and A2780 cell lines.CSPG4 is mainly located on the cell membrane in different epithelial ovarian cancer cell lines.3.MTT showed the growth of HEY sh RNA-CSPG4 cells was significantly inhibited compared with HEY sh RNA-control,and the difference between them was statistically significant(P<0.05).4.Soft agar assay showed the number of colonies formed in the sh RNA control group was significantly greater than that in the ovarian cancer HEY cell line transfected with sh RNA-CSPG4.The difference in colony formation between the two groups was statistically significant(P<0.01).5.Wound healing showed after 24 hours,the scratches of HEY sh RNA-control cells had completely healed,while there were still significant gaps in HEY sh RNA-CSPG4 cells,and there was a significant difference between the two(P<0.01).6.The protein expression of p-c-Met,p-ERK1/2 and ERK1/2 in sh RNA-CSPG4 transfected HEY cells was weaker than that in the sh RNA-control control group,but there was no difference in protein expression of p-Akt between the two groups.7.In ovarian cancer HEY cells,c-Met expression was decreased by sh RNA,and the expression of c-Met,p-c-Met and p-ERK1/2 proteins was weaker than that in the control group,but the expression of p-Akt and Akt proteins was not significantly changed.Conclusions:1.Inhibition of CSPG4 expression can inhibit the proliferation and migration of epithelial ovarian cancer cells.2.CSPG4 may promote the development of malignant epithelial ovarian cancer through the c-Met and ERK1/2 signaling pathways.