Study Of The Function And Mechanism Of TRIP13 In Colorectal Cancer Progression

Objectives:(1)To detect the expression of TRIP13 in colorectal cancer and to investigate the relationship between TRIP13 and the prognosis of colorectal cancer patients.(2)To investigate the effect of TRIP13 on the biological function of colorectal cancer cells.(3)To investigate the mechanism of TRIP13 in the progression of colorectal cancer.Methods:(1)The expression of TRIP13 in colorectal cancer tissues was detected by Western Blotting and immunohistochemistry,the relationship between TRIP13 expression and clinicopathological factors as well as prognosis was analyzed.(2)TRIP13 was knocked down and overexpressed in colorectal cancer cell lines,and MTT,clone formation as well as Transwell invasion and migration assays were performed to detect the effect of TRIP13 expression on the proliferation,invasion and migration.Subcutaneous tumorigenesis experiment with TRIP13 knockdown and overexpression colorectal cancer cells was used to compare the effect of TRIP13 in tumorigenicity.(3)The effect of TRIP13 knockdown and overexpression on the expression of E-cadherin,N-cadherin,β-catenin and snail was detected by Western Blotting,to determine whether TRIP13 was involved in the EMT in invasion and migration of colorectal cancer cells.Immunoprecipitation and MS were applied to identify TRIP13 interacting proteins.The target protein was knocked down in TRIP13-overexpressed cells and the expression of E-cadherin,N-cadherin,β-catenin and snail was detected by Western Blotting.To detect the expression of targeted protein in colorectal cancer tissues,correlation analysis was adopted to clarify the relationship of TRIP13 and the target protein.Results:(1)The expression of TRIP13 in colorectal cancer tissues was significantly higher than that in normal intestinal epithelium tissues,TRIP13 was closely related to tumor stage(pTNM),CEA and CA19-9.Patients with high expression of TRIP13 had a worse prognosis.(2)Knockdown of TRIP13 significantly inhibited the proliferation,invasion and migration ability of colorectal cancer cells,while overexpression of TRIP13 inversed the phenomena.Tumorigenicity experiments in nude mice also confirmed that the ability of tumorigenesis was weakened after knockdown of TRIP13,while the ability of tumorigenesis increased after TRIP13 was overexpressed.(3)After knockdown of TRIP13,the expression of E-cadherin increased,while the expression of N-cadherin,β-catenin and snail decreased.However,the expression of E-cadherin decreased and the expression of N-cadherin,β-catenin and snail increased after TRIP13 overexpressed.Mass spectrometry results showed that TRIP13 interacts with YWHAZ;knockdown of YWHAZ in TRIP13 overexpression colorectal cancer cells promotes the expression of E-cadherin,and inhibits the expression of N-cadherin,beta-catenin and snail.Spearman correlation analysis showed that there was a positive correlation between YWHAZ and TRIP13.Conclusion:(1)TRIP13 is highly expressed in colorectal cancer tissues,and TRIP13 is an independent risk factor for the prognosis of colorectal cancer.(2)TRIP13 promotes the proliferation,invasion,migration and tumorigenicity ability of colorectal cancer cell.(3)TRIP13 plays a role in regulating EMT by targeting YWHAZ to promote invasion and metastasis of colorectal cancer.