Expression Of NET-1 In Colorectal Adenocarcinoma Tissues And Clinical Significance

Objective: We sought to investigate the roles and the potential mechanisms of NET-1 in the occurrence and progression of colorectal cancer.By this study,we want to provide a promising biomarker of colorectal cancer.Methods: NET-1 mRNA and protein expression levels in tumor tissues and its adjecent non-tumor tissues from 120 CRC patients was detected by RT-PCR and immunohistochemistry(IHC).The relationship between NET-1 protein expression level and clinical parameters of CRC tissues was analyzed with SPSS software 19.0.NET-1 cDNA CDS fragments were subcloned into pIRES2-EGFP eukaryotic expression vector after verification by DNA sequencing.pIRES2-EGFP/NET-1 or control vectors were transfected into HCT116 CRC cells.The sTablele expression cell lines were obtained through G418 selection.The change of cell growth and invasion were detected in CRC cells by cell counting assay and cell invasion assay after transfection.Moreover,protein level of MMP-9 was detected by Western-blot.Knockdown NET-1 in CRC cells was performed via shRNA.Cell growth;and migration were detected in shNET-1 cells by cell count assay and migration assays.Proteins level of MMP-9 was also detected by Western-blot after knockdown.Follow-up: Patients were divided into NET-1 positive and negative group and followed up.Data were collected for survival regression analysis by SPSS19.0.Results:The positive rate of NET-1 expression in colorectal cancer tissue was higher than that in normal population;the positive expression of Ki-67,MMP-9,and VEGF were positively correlated to the expression of NET-1.The relative expression level of NET-1 mRNA in normal tissues and peritumor tissues were lower than that in colorectal cancer tissues.Western-blot: Consistently,the expression of NET-1 protein in the normal colon epithelium and peritumor tissue were lower than that in colorectal cancer tissue.Survival analysis: Survival analysis showed that there was significant difference in the survival time of NET-1 positive group and the negative group(P<0.05).Conclusion:1.The expression of NET-1 plays an important role in the occurrence and development of colorectal cancer.2.The expression of NET-1 in colorectal cancer has the close relation with the degree of differentiation,TNM staging and lymph node metastasis and has significant correlation with the expression of Ki-67,MMP-9,and VEGF.3.The expression of NET-1 in colorectal cancer is related to the survival of patients,and the prognosis with high expression of NET-1 is poorer.