Cadherin 12 Promote Angiogenesis Through PI3K-Akt Signaling Pathway In Colorectal Cancer

OBJECTIVE: To study the expression of cadherin 12(CDH-12)in colon cancer cell line and colorectal cancer tissue.To explore the effect of CDH-12 on neovascularization of colon cancer cells and its mechanism.Methods: Immunohistochemical technique was used to detect the expression of CDH-12 in colorectal cancer tissues.The correlation between CDH12 and platelet endothelial cell adhesion molecule-1(CD31)in colorectal cancer tissues was detected by immunohistochemistry.Western blotting was used to screen CDH-12 overexpressing cell lines in 9 colon cancer cell lines.High expression cell line HCT116 was transfected with SiRNA and Western blotting was used to detect the interference effect.Endothelial tube formation assay was performed to detect the effect of CDH12 on angiogenesis.Changes of Signaling Pathway was detected by Western blot.Enhance CDH12-expression in HT-29 cell line,and its effect on umbilical vein endothelial cells was detected by tubule formation experiment.Western blotting was used to detect the changes of signal pathway in HT-29 cells.The Akt phosphorylation inhibitor AZD5363 was used to the CDH-12-overexpressed HT-29 cells and then Implement the small tube formation experiment.AT the same time Akt phosphorylation level was detected.Results: The expression of CDH12 was positively correlated with CD31 according to the results of continuous immunohistochemical staining.Immunofluorescence showed the area CDH12 highlight,CD31 was also highlighted.The expression of CDH-12 in sw480,sw48,sw620,HCT116 and NCIH716 cells was significantly higher than that in other cell lines.After successful Knock down of CDH-12 in HCT116 cells,the angiogenesis ability of HCT116 colon cancer cell line was significantly lower and the phosphorylation of Akt was significantly decreased than that of control group.After successful overexpression of CDH-12 in HT-29 cells,the angiogenesis ability of HT-29 colon cancer cell line was significantly hingher and the phosphorylation of AKT was significantly increased than that of control group.Phosphorylation inhibitor AZD5363 was used to treat CDH-12-overexpressing HT-29 cells,and the ability of angiogenesis of umbilical vein endothelial cells was reduced and the degree of phosphorylation of AKT was decreased.Conclusion: The expression of CDH12 in colorectal cancer tissues was positively correlated with vascular marker CD31.Indicating that CDH12 is positively correlated with angiogenesis in colorectal cancer.Knock down CDH-12 in HCT116 and the vascular formation was weakened indicating CDH12 promote tumor angiogenesis.After knocking down of CDH-12 in HCT116,Akt phosphorylation was reduced.Akt phosphorylation was significantly enhanced in CDH-12-overexpressing HT-29 cells and angiogenesis was enhanced compared with control groups.Phosphorylation inhibitor was used to treat CDH-12-overexpressing HT-29 cells and the angiogenesis ability of HT-29 and the phosphorylation of Akt was recovered indicating that Cadherin 12 promote angiogenesis through PI3K-Akt signaling pathway in colorectal cancer.