Effect And Mechanism Of ITGA5 On Metastasis And Prognosis Of Colorectal Cancer

Background Colorectal cancer(CRC)is one of the common human intestinal malignancies in China and in the world,which ranks third among men and second among women.At present,the main treatment methods for CRC include surgical excision,chemoradiotherapy combined with targeted gene therapy or without and etc.Although big processes have been achieved in the diagnostic and treatment methods,the 5-year survival rate for CRC patients remains low due to its stealthiness and high metastasis rate.Therefore,it is of importance to further elucidate the molecular mechanisms underlying the occurrence and development of CRC,and find the potential therapeutic targets to improve the treatment effect and prognosis of CRC.Research purposes The objective of this study is to explore the relationship between ITGA5 expression levels and the clinicopathological features and prognosis of CRC patients.At the same time,the biological function of ITGA5 in colon cancer progression was explored by using in vitro and in vivo experiments to further clarify the underlying mechanisms of ITGA5 in cell proliferation,invasion,apoptosis and metastasis in colon cancer.Research methods1.Second-generation sequencing technology was used to screen the differentially expressed genes between colon cancer tissues and the adjacent normal tissues,and RT-PCR technology was used to test and verify the differential expression levels of ITGA5 in colon cancer tissues and the adjacent normal tissues.2.Retrospectively analyze the correlation between the expression level of ITGA5 and the overall survival and clinicopathological characteristics of colon cancer patients,and clarify the clinical value of the expression levels of ITGA5 in colon cancer.3.The effects of ITGA5 on the prolifetaion,apoptosis,invasion migration and tumorigenesis of colon cancer cells were analyzed by the means of CCK-8,flow cytometry,transwell cell invasion,wound healing assay and xenograft tumor models.4.The relationship between ITGA5 and O-Glc NAc was analyzed by co-immunoprecipitation and O-Glc NAcylation enzyme labeling experiments,and the effect of O-Glc NAcylation on the stability of ITGA5 protein was analyzed by western blotting technique.Research results1.The results of next-generation sequencing showed that there were multiple differentially expressed signal molecules between the colon cancer tissues and the adjacent non-tumor tissues,among which ITGA5 had significant differences,which was confirmed by RT-PCR technology.2.Retrospective analysis showed that the expression level of ITGA5 was positively correlated with the TNM grade,high incidence of lymphoid tissue metastasis,and poor prognosis in patients with colon cancer.3.Overexpression of ITGA5 promoted the proliferation,migration,invasion and in vivo tumor formation of colon cancer cells and inhibited apoptosis,while ITGA5 downregulation inhibited cell proliferation,migration,invasion and in vivo tumor formation and promote cell apoptosis.4.In colorectal cancer cells,ITGA5 protein could be O-Glc NAcylated,which enhanced the stability of ITGA5 protein.Conclusion This study demonstrates that ITGA5 is upregulated in colon cancer tissues and asscociated with tumor metastasis and prognosis.ITGA5 protein could be O-Glc NAcylated,which enhanced the stability of ITGA5 protein.This study indicates that ITGA5 can be used as a new potential prognostic biomarker and therapeutic target for colon cancer,which have certain research significance.