钙粘素12促进结直肠癌中新生血管的形成

摘要

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.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.The colon cancer cells transfected with siRNA fragments with meaningless sequence were negative control group, and colon cancer cells with only LipofectamineTM 2000 transfected medium were blank control group.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 endothelial tube formation assay was performed to detect the effect of CDH12 on angiogenesis.Enhance CDH12-expression in HT-29 cell line, and its effect on human umbilical vein endothelial cells was detected by tubule formation experiment.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.After successful Knock down of CDH-12 in HCT116 cells, the angiogenesis ability of HCT116 colon cancer cell line was significantly lower 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 than that of control group.Conclusion Cadherin 12 promotes angiogenesis in colorectal cancer.%目的 研究钙黏蛋白12(CDH-12)在结直肠癌组织与结直肠癌细胞株中的表达及其对结肠癌新生血管的影响.方法 免疫组化连续切片,免疫荧光,检测结直肠癌组织中CDH12和新生血管密度的相关性,CD31作为新生血管内皮细胞标志物,即用CD31反映新生血管的位置及密度.将转染带有无意义序列的siRNA片段的结肠癌细胞组为阴性对照组,将仅加入LipofectamineTM 2000 转染介质的结肠癌细胞组为空白对照组.应用Western免疫印迹法在9株结肠癌细胞株中筛选CDH-12高表达细胞株;siRNA瞬转干扰高表达CDH12的细胞株HCT116,小管形成试验检测敲低CDH12后肠癌细胞株HCT116对人脐静脉内皮细胞成管的影响.过表达HT-29细胞系的CDH-12,通过小管形成实验,检测其对人脐静脉内皮细胞成管的影响.结果 根据CDH12染色结果评分和CD31所反映的微血管计数,做相关分析,结果表明CDH12表达与新生微血管数呈正相关.免疫荧光显示代表CDH12的高亮区,CD31也高亮,反映新生血管集中区域与CDH12高表达区域相一致.下调HCT116细胞中CDH-12表达之后,培养过该组癌细胞的培养液培养人脐静脉内皮细胞小管形成能力较对照组明显下降;过表达HT-29细胞系的CDH-12后,使得人脐静脉内皮细胞成管能力增强.结论 CDH12在结直肠癌细胞系及结直肠癌组织中起到促进新生血管形成的作用.