Hsa＿circ＿0060967 As CeRNA Regulates MiR-338-3p Affects The Invasion And Metastasis Of Colorectal Cancer Cells

AIMS: Through detected the expression of hsa＿circ＿0060967(circ RAE1)in colorectal cancer tissues and cells,to analyze its expression level and clinical pathologic features and prognosis of correlation,and then through external intervention hsa＿circ＿0060967expression,testing its impact on the behavior of colorectal cancer cell invasion and metastasis.Finally,research the relationship between hsa＿circ＿0060967,mi R-338-3p and TYRO3,confirmed its function effect of molecules,so as to provide a new clinical diagnosis for the treatment of colorectal cancer or therapeutic targets.METHODS:To collect the information of colon cancer,paracancer tissues and clinicopathology,the expression levels of hsa＿circ＿0060967 in colorectal cancer tissues,paracancer tissues and cell lines were detected by q RT-PCR,and the correlation between hsa＿circ＿0060967 and clinicopathological characteristics was analyzed by spss22.0.To further study the stability and localization of hsa＿circ＿0060967 in cells at the cell level,the expression levels of EMT marker genes were detected by cell score assay,Transwell assay,q RT-PCR and WB,and evaluated the effects of transfection and co-transformation of hsa＿circ＿0060967,hsa＿circ＿0060967/ mi R-338-3p,hsa＿circ＿0060967/TYRO3 on the biological functions of colon cancer cells(epithelial mesenchymal transformation,metastasis,migration and invasion).q PCR was used to detect the expression levels of mi R-338-3p in cancerous tissues and para-cancerous tissues as well as in cancer cells,and verified the targeting relationship between hsa＿circ＿0060967/ mi R-338-3p and mi R-338-3p /TYRO3 by double luciferase reporter.RESULT:In this study,it was found that hsa＿circ＿0060967 expression in CRC tissues and cells was more stable and significantly up-regulated than that in para-carcinoma tissues and normal colonic epithelial cells,and its high expression level in CRC tissues was positively correlated with CRC tumor size(>5cm),lymphatic metastasis and TNM(III、IV 期)stage.The results of fluorescence in situ hybridization showed that hsa＿circ＿0060967 and mi R-338-3p colocated in cytoplasm.In CRC cells,after interfering with hsa＿circ＿0060967,the migration and invasion of CRC cells decreased,the expression of E-cadherin increased,the expression of Vimentin decreased,and the expression of mi R-338-3p increased compared with that of the control group.Co-transfected Hsa＿circ＿0060967 and mi R-338-3p into CRC cells(SW620 and HT29),compared with the negative control group(LV-NC+mi R-NC),interference with hsa＿circ＿0060967 resulted in increased expression of mi R-338-3p but the expression of TYRO3 and cell invasion and migration were decreased.There was no change in the expression of hsa＿circ＿0060967 by interfering with mi R-338-3p,but the invasion and migration ability of CRC cells and the expression of TYRO3 were increased.However,the invasion and migration ability of the cells decreased when only interfering with TYRO3 than when only interfering with mi R-338-3p.CONCLUSION: Hsa＿circ＿0060967 plays the role of sponge in absorbing a large amount of mi R-338-3p,which weakens the regulatory capacity of mi R-338-3p on its targeted protein TYRO3,so that TYRO3 plays a role in promoting the progression of colon cancer.It suggests that hsa＿circ＿0060967 may be a potential molecular target for the development of anticancer drugs.