Expression Of EphA7 In Osteosarcoma And Its Biological Effect On MG-63 Cells

Background and purpose: EphA7 is a membrane protein that plays a key role in the normal functional activities of the body.The place where it often plays a role is the cell membrane,but what role it plays in osteosarcoma has not been studied.Therefore,in this experiment,the content of EphA7 in osteosarcoma tissues of45 patients was determined,and related methods were used to investigate whether EphA7 had an effect on some biological effects of human osteosarcoma MG-63 cells.In order to understand the role of EphA7 protein in osteosarcoma.Methods: The osteosarcoma tissue and corresponding normal adjacent tissues of45 osteosarcoma patients were collected and stored in liquid nitrogen.Culture human osteosarcoma MG-63 cells,transfect human osteosarcoma MG-63 cells according to siRNA Reagent System kit instructions,and divide them into siRNA-EphA7 group(EphA7-siRNA transfection),siRNA-Control group(negative control siRNA transfection)and blank group(untransfected).Real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR)was used to determine the mRNA expression of EphA7 in tissues,and Western blot was used to determine the protein expression of EphA7 in cells.Cell counting kit(CCK-8),cell scratch test,and flow cytometry were used to determine the cell's proliferation activity,cell migration ability,and cell apoptosis.Results: The results of RTFQ-PCR and Western blot showed that the expression levels of EphA7 mRNA and protein in osteosarcoma tissue were significantly higher than those in normal adjacent tissues(P <0.05).Compared with the blank group and siRNA-Control group,siRNA-In the EphA7 group,the expression of EphA7 mRNA and protein level were significantly reduced(P <0.05).Compared with the blank group and the siRNA-Control group,the mRNA expression and protein level of EphA7 did not change significantly(P> 0.05).CCK-8 measurement results showed that the absorbance(D)value of the siRNA-EphA7 group was significantly lower than that of the blank group and the siRNA-Control group(P <0.05).There was no significant change in the absorbance(D)value of the blank group and thesiRNA-Control group(P> 0.05).Cell scratch experiments showed that compared with the blank group and the siRNA-Control group,the healing rate of the siRNA-EphA7 group was significantly reduced after scratching(P <0.05).There was no significant change in the healing rate(P> 0.05).Flow cytometry showed that compared with the blank group and the siRNA-Control group,the apoptosis rate of the siRNA-EphA7 group increased significantly(P <0.05).Compared with the blank group and the siRNA-Control group,there was no significant change in the apoptosis rate of the two(P> 0.05).Conclusion: EphA7 is very low in normal organisms,but it is significantly higher in osteosarcoma patients.Down-regulating the expression of EphA7 in human osteosarcoma MG-63 cells can effectively limit the proliferation and migration of human osteosarcoma MG-63 cells and promote their apoptosis.