Mechanism Of Brexpiprazole In Inhibiting Colon Cancer And Increasing The Sensitivity Of Cetuximab Treatment

Objective:To investigate the inhibitory effects of brexpiprazole on human colon cancer cells and the potential mechanism of increasingthe sensitivity of colon cancer cells to the cetuximab.Methods:In vitro study,CCK-8 was used to detect and compare the effects of brexpiprazole on the proliferation activity of human colon cancer cells HCT116,SW620 and normal human colon epithelial cells NCM460.Would healing,transwell,and flow cytometry were adopted to identify the effects of brexpiprazole on the migration and apoptosis in human colon cancer cell lines HCT116 and SW620.Western blot was performed to detect the changes of expression level in apoptotic and EGFR signaling pathway related proteins.In HCT116 cells,CCK-8 was utilized to confirm the inhibitory effect of brexpiprazole in combination with cetuximab on the proliferation.Western blot was adopted to examine the changes in the expression of EGFR signaling pathway proteins in colon cancer cells after combined treatment.In vivo study,the nude mouse subcutaneous xenograft tumor model was established by HCT116 and the tumor growth curve was drawn.IHC was conducted to testify the expression of survivn protein in the tumor tissues of HCT116 xenografts,and western blot was executed to detect the effects of brepiprazole alone or in combination with cetuximab on the expression of EGFR-related proteins in tumor tissues of HCT116 xenograft mice model.Results:①Brexpiprazole has a selective inhibitory effect on the proliferation of NCM460,HCT116 and SW620 cells respectively.②Brexpiprazole can significantly inhibit the migration of HCT116 and SW620 cells in vitro and induce apoptosis in a dose-dependent manner.③Brexpiprazole down-regulated the expression of survivin,caspase,9,p-AKT,p-PI3K P85,p-P38 MAPK,p-ERK1/2,p-Raf protein in colon cancer cells in a dose-dependent manner.④Brexpiprazole can remarkably suppress tumor proliferation in vivo,and down-regulated the expression levels of survivin,p-AKT,p-PI3K P85,p-P38 MAPK,p-ERK1/2,p-Raf protein in cancer tissues.⑤Compared with the control group and the single medication group,brexpiprazole combined with cetuximab has a prominent inhibitory effect on the proliferation of HCT116 cells.⑥The results of nude mice tumor-bearing experiments indicated that brexpiprazole combined with cetuximab in tumor volume is noteworthy than that of in the single medication group.Compared with the control group and the single medication group,the combination of brexpiprazole and cetuximab could markedly down-regulated the expression of survivin,p-AKT,p-PI3K P85,p-P38 MAPK,p-ERK1/2,and p-Raf proteins in tumor tissues.Conclusion:Brexpiprazole can inhibit the proliferation,migration and induce apoptosis of human colon cancer cells.The combined application of brexpiprazole and cetuximab can produce a notable synergistic anti-tumor effects and increase the sensitivities of colon cancer cells to targeted therapy.The potential mechanism for anti-tumor biological activities of brexpiprazole in vivo and in vitro might be related to the refraining of PI3K-AKT and MAPK-ERK signaling pathways of EGFR.