TRIP6 Regulates The Proliferation,Migration,Invasion And Apoptosis Of Osteosarcoma Cells By Activating The NF-?B Signaling Pathway

Objective: Osteosarcoma(Os)is one of the most common primary malignant bone tumors in pediatrics and adolescents,and it is a rare mesenchymal tumor that occurs mainly in the metaphysis of long diaphysis,especially in the distal femur,proximal tibia,and humerus.Os accounts for about 20% of primary malignant bone tumors and 0.5% of malignant tumors.Os mostly occurs in epiphyseal region,which is rich in blood supply.The incidence of hematogenous metastasis is high,occurs early and progresses quickly.Previous studies have shown that if untreated,80% of patients with Os develop lung metastases,which are often fatal.Currently,drugs used to treat Os include adriamycin,ifosfamide,methotrexate and cisplatin.Some patients have poor prognosis or drug resistance after chemotherapy,which is difficult to improve despite different treatment combinations.In addition,high doses of chemotherapy drugs have side effects.Therefore,the in-depth study of the molecular mechanism of the occurrence and development of Os and the search for the target of gene therapy for Os will provide new ideas and experimental basis for the effective treatment of Os.The purpose of this study was to investigate the effects of TRIP6 on proliferation,apoptosis,invasion and migration of Os cells.Methods: U2 OS and MG63 Os cell lines were cultured with fetal bovine serum and high glucose medium.Four human plasmids were constructed,namely sh RNA-TRIP6 plasmids,sh NC plasmids,pcDNA Flag TRIP6 plasmids and pex-1 plasmids.The sh RNA-TRIP6 plasmid group was the silent TRIP6 group,the pcDNA Flag TRIP6 plasmid group was the TRIP6 overexpression group,the sh NC plasmid group was the control group of the silent TRIP6 group,and the pex-1 plasmid group was the control group of the TRIP6 overexpression group.The expression of TRIP6 in osteosarcoma cell lines U2 OS and MG63 was detected by Western blot.The proliferation ability of cells in each group was determined by MTT assay.The migration ability and invasion ability of cells in each group were detected by Transwell migration test and invasion test.Cell apoptosis in each group was detected by flow apoptosis kit.Results: after transfection of human Os cell lines U2 OS and MG63 into plasmids of various experimental and control groups,the results of WB experiment showed that: for the control group transfected with pex-1 plasmids,the expression level of TRIP6 was significantly increased in the experimental group transfected with pcDNA Flag TRIP6 plasmids.For the sh NC plasmid transfected control group,TRIP6 expression was significantly reduced in the sh RNA-TRIP6 plasmid transfected group.The statistical difference between the two groups was statistically significant(P<0.05).MTT test results showed that compared with the control group,the proliferation of U2 OS and MG63 cells transfected with pcDNA Flag TRIP6 plasmid was significantly enhanced,while the proliferation of U2 OS and MG63 cells transfected with shrna-trip6 plasmid was significantly decreased,with statistically significant differences between the two groups(P<0.05).Transwell migration test and invasion test results showed that U2 OS and MG63 cells transfected with pcDNA Flag TRIP6 plasmid had significantly increased migration and invasion ability compared with the control group,while U2 OS and MG63 cells transfected with shrna-trip6 plasmid had significantly decreased migration and invasion ability,with statistically significant differences between the two groups(P<0.05).The apoptosis rate of U2 OS and MG63 cells transfected with pcDNA Flag TRIP6 plasmid was significantly decreased compared with the control group,while the apoptosis rate of U2 OS and MG63 cells transfected with sh RNA-TRIP6 plasmid was significantly increased,with statistically significant differences between the two groups(P<0.05).Conclusion: in this study,we successfully transfected the shrna-trip6 plasmid and pc DNA Flag TRIP6 plasmid into human Os cell lines U2 OS and MG63 cell lines.The results of MTT assay,Transwell migration assay,invasion assay and flow apoptotic cell assay showed that TRIP6 promoted the proliferation,migration and invasion ability of Os cells U2 OS and MG63,and reduced the apoptosis rate.This study showed that TRIP6 gene could be a potential target for diagnosis and treatment of Os,laying a foundation for further exploration of the role of TRIP6 in the pathogenesis of Os and providing a theoretical basis for targeting TRIP6 in the treatment of Os.