Effects On Proteasome Inhibitor Marizomib Sensitizing Cisplatin-induced Cytotoxicity And Apoptosis In Cervical Cancer

Objectives:To elucidate the effect of marizomib combined with cisplatin on apoptosis,proliferation,molecular mechanism and potential clinical significance of cervical cancer cells and their xenografts.Methods:The effects of marizomib or in combination with cisplatin on different types of cervical cancer cell lines were detected by CCK-8 and clonogenic assay.According to the IC₅₀value of marizomib on cervical cancer cells,different drug concentrations were screened or used in combination with cisplatin.Western blot technology and flow cytometry were used to detect the apoptosis effects of cisplatin and marizomib alone and in combination on cervical cancer cells.Cytokine antibody chip experiments were used to detect the expression changes of multiple cytokines after drug treatment.After bioinformatics big data analysis,Western blot was used to detect the changes in the expression of different molecules,such as cell proliferation and apoptosis.Construct xenograft models of nude mice to observe the difference in tumor volume in each group after drug treatment;observe tumor hemorrhage,necrosis changes and the invasion of cancer cells to surrounding tissues in vivo after drug treatment by H&E;use immunohistochemical techniques to detect cervical cancer Localization and expression of key proteins in cancer xenografts.Western blot was used to detect the expression of apoptosis-related proteins in subcutaneous transplanted tumors of nude mice and the expression of differential signaling molecules analyzed by cytokine antibody chip.The online database data was used tomine and analyze the m RNA expression levels,prognosis and correlation of key genes in cervical cancer tissues.Results:（1）Compared with the immortalized cervical epithelial cells H8,marizomib significantly inhibited the proliferation of cervical cancer cells and promoted the cleavage of the apoptotic factors PARP and caspase 3（P<0.05）,and the effect was more significant when combined with cisplatin（P<0.05）.（2）Marizomib combined with cisplatin significantly inhibited the growth and invasion of xenograft models（P<0.05）.In terms of molecular mechanism,up-regulated the expression of Ang-1and down-regulated the expression of Flt-3L,SCF and Tie-2 signaling are involved in marizomib enhancing the cytotoxic effects of cisplatin on cervical cancer（P<0.05）;TCGA online database analysis results showed that in 305 cervical cancer samples,the expression levels of Ang-1,Tie-2,Flt-3L and SCF m RNA were higher,while the expression level of Ang-1 was relatively low.Meanwhile,patients with lower Ang-1 m RNA expression level had poorer prognosis,which was positively correlated with Tie-2（P<0.05）.Conclusion:（1）Marizomib induces cervical cancer cell apoptosis,inhibits proliferation,and sensitizes the effect of cisplatin therapy on cervical cancer cells;（2）Up-regulation of Ang-1 and down-regulation of Flt-3L,SCF and Tie-2 signaling are involved in marizomib sensitizing cisplatin in the treatment of cervical cancer cells.