The Expression Of RIP3 And Its Function Study In Colorectal Cancer

ObjectivesThe aim of this study is to discuss the mRNA and protein expression of the Receptor-interacting protein kinase 3(RIP3)in colorectal cancer tissues and its function in colorectal cancer cells,which contribute to reveal the molecular mechanisms for incidence and development of colorectal cancer.Methods1.Quantitative PCR and Western blot analysis were used to examine RIP3 level of mRNA and protein in primary colorectal cancer and the normal colonic mucosa respectively.2.RIP3 clinicopathological significance was assessed by immunohistochemical staining in primary colorectal cancer paired with noncancerous tissues.Kaplan-Meier and Cox proportional hazards model analysis were performed to assess the correlation between RIP3 expression and patients OS rate and DFS rate.3.The biological function of RIP3 overexpression was measured by CCK8 assay,plate colony formation assay and transwell assay.Fluorescence detection has been used to detect cell apoptosis.Results1.The results of Real-time PCR suggest us that 60.5%(23/38)colorectal cancers showed at least a 2-fold decrease in RIP3 mRNA levels compared with that of the adjacent normal tissue among the 38 paired specimens.Western blot confirmed that samples expressed a lower level of RIP3 protein in colorectal cancerous tissues than that in paired normal mucous.2.Immunohistochemistry indicated that RIP3 expression was significantly associated with T stage(P=0.027),M stage(P=0.016)and AJCC stage(P=0.002).Kaplan-Meier and Cox proportional hazards model analysis showed the patients with negative RIP3 expression had a lower OS rate and DFS rate than patients with positive expression(P=0.004,P=0.015,respectively).3.Overexpression of RIP3 significantly suppressed the proliferation,colony formation,migration,invasion and caused pronounced apoptosis of colorectal cancer cells.Conclusions1.The expression of RIP3 gene in colorectal cancerous tissue is apparently lower compared with paired normal tissues.2.RIP3 gene was associated with some disadvantageous clinicopathologic factors,including T stage,N stage,AJCC stage and was a predictor of survival in colorectal cancer.3.These results suggest that RIP3 exhibits a suppressive role in the incidence and development of colorectal cancer.Our study provides a new therapeutic target for colorectal cancer treatment.