| BackgroundCurrent research shows that ADAM 17 is highly expressed in malignant tumors including gastric cancer,breast cancer,and lung cancer,and its expression is related to tumor pathology,chemotherapy drug response,etc.,but the expression of ADAM 17 in colorectal cancer is less studied.The correlation and mechanism of expression and tube formation of lymphatic endothelial cells are still unclear.PurposeClarify the expression of ADAM17mRNA and AD AM17 protein in colorectal cancer and normal tissues,analyze the correlation between the expression of ADAM17mRNA and ADAM 17 protein and tumor pathology,and explore the effect of ADAM 17 expression on lymphatic endothelial cell tube formation.Methods(1)Real time PCR method was used to detect the expression of ADAM17mRNA in fresh colorectal cancer tissues and matched normal tissues;analyze the correlation between the expression of ADAM17mRNA in tumor tissues and tumor stage;(2)Immunohistochemical method was used to detect the expression of AD AM17 protein in colorectal cancer tissues and matched normal tissues;immunohistochemical method was used to analyze the correlation between the expression of AD AM17 protein in tumor tissues and various pathological indicators of tumors;(3)Western blot method was used to detect the expression of ADAM 17 protein in normal cell lines and 6 colorectal cancer cell lines,and to select the cell line with the highest expression of AD AM17 protein;(4)Real time PCR method is used to detect the siRNA with the best interference effect of siRNA-391,siRNA-1356 and siRNA-483;(5)The selected colorectal cancer cell lines were divided into a control group,a plasmid overexpression group,a negative control group,and an interference group,and the cell growth curves of each group were drawn;the cell absorbance(A)value of each group was detected by the MTT method;(6)Co-culture the selected cell lines with HLEC human lymphatic endothelial cells and divide them:into four groups:co-culture control group,co-culture+overexpression plasmid group,co-culture+ interference group,and co-culture+negative control group.Observe the lymphatic endothelial cell tube formation.Results(1)The expression of ADAM 17 mRNA in cancer tissues of 36 patients with colorectal cancer was significantly higher than that of matched normal tissues(p<0.001);the expression of AD AM17 protein in cancer tissues of 100 patients with colorectal cancer was significantly higher than that of matched normal tissues Organization(p<0.001);(2)Among 36 patients,the expression level of ADAM17 mRNA in colorectal cancer tissues of T3+T4 group was significantly higher than that of T1+T2 group(p=0.002),and AD AM17 in colorectal cancer tissue of patients with lymph node metastasis group The mRNA expression level was significantly higher than that in patients without lymph node metastasis(p<0.001);(3)In 100 cases of colorectal cancer,the expression of AD AM17 protein was correlated with the degree of tumor differentiation,T stage,N stage,pTNM stage,nerve invasion,vascular tumor thrombus,and vein invasion;(4)Western blot method was used to detect the expression of ADAM 17 in different colorectal cancer cell lines and normal intestinal mucosal FHC cell lines.The results showed that the expression of AD AM17 was the highest in Caco2 cell lines;(5)Real time PCR showed that among siRNA-391,siRNA-1356,and siRNA-483.siRNA-391 had the best interference effect;(6)The growth rate of Caco2 cells overexpressed by ADAM 17 is significantly accelerated,especially after entering the logarithmic growth phase;(7)The absorbance(A)value of the cells in each group was detected by the MTT method.The results showed that the absorbance value of the cells in the overexpression plasmid group increased faster;(8)Caco2 cell line was co-cultured with HLEC human lymphatic endothelial cells.Observe the formation of lymphatic endothelial cells in the co-culture control group,co-culture+overexpression plasmid group,co-culture+interference group and co-culture+negative control group.+The rate of lymphatic endothelial cell tube formation in the overexpression plasmid group was significantly higher than that in the control group and the negative control group,and no lymphatic tube formation was observed in the interference group.Conclusions(1)The expression levels of ADAM 17 mRNA and ADAM 17 protein in cancer tissues of patients with colorectal cancer are significantly higher than those of matched normal tissues;(2)The expression level of AD AM 17 mRNA in colorectal cancer tissue is correlated with the T and N stages of the tumor;(3)The expression of ADAM 17 protein in colorectal cancer tissue is correlated with the degree of tumor differentiation,T stage,N stage,pTNM stage,nerve invasion,vascular tumor thrombus,and vein invasion;(4)ADAM 17 overexpression promotes the proliferation of Caco2 cells;(5)Co-culture of Caco2 human colon cancer and HLEC human lymphatic endothelial cells.ADAM 17 overexpression is beneficial to the formation of lymphatic endothelial cells.Conversely,interference with ADAM 17 expression inhibits the formation of lymphatic endothelial cells. |
PDF Full Text Request