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目的探讨Aurora-A高表达对食管鳞癌血管生成及血管内皮生长因子受体-2(vascular endothelial growthfactor receptor 2,VEGFR-2)表达的影响。方法将绿色荧光蛋白(green fluorescent protein,GFP)标记的Aurora-A全长表达质粒pEGFP-C1-Aurora-A转染至食管鳞癌细胞KYSE150,经G418筛选获得Aurora-A高表达的细胞株(Aurora-A高表达组),同时设空质粒pEGFP-C1转染组(阴性对照组)和未转染组作为对照。Western blot检测各组细胞中Aurora-A蛋白的表达;采用小管形成及鸡胚尿囊膜试验检测Aurora-A高表达对肿瘤血管生成的影响;免疫组化法检测Aurora-A高表达对裸鼠移植瘤中微血管密度(microvessel density,MVD)的影响;Western blot检测Aurora-A高表达对KYSE150细胞中VEGFR-2及p-VEGFR-2(Tyr1059)表达的影响。结果 Aurora-A高表达组中总Aurora-A蛋白的表达量约为阴性对照组和未转染组的2.5倍,提示Aurora-A高表达细胞系构建成功;Aurora-A高表达组生成的血管数量、MVD值和p-VEGFR-2的表达水平均显著高于阴性对照组(P<0.01)。结论 Aurora-A高表达可促进食管鳞癌中的血管生成,其机制可能与活化VEGFR-2有关。
Objective To investigate the effect of Aurora-A overexpression on the angiogenesis and the expression of vascular endothelial growth factor receptor 2 (VEGFR-2) in esophageal squamous cell carcinoma. Methods The full-length Aurora-A expression plasmid pEGFP-C1-Aurora-A labeled with green fluorescent protein (GFP) was transfected into KYSE150 esophageal squamous cell carcinoma cell line, and the cell line Aurora-A highly expressed Aurora-A high expression group). At the same time, empty plasmid pEGFP-C1 transfection group (negative control group) and untransfected group as control. The expression of Aurora-A protein in each group was detected by Western blot. The effect of Aurora-A expression on tumor angiogenesis was detected by tubule formation and chick embryo allantoic membrane assay. Immunohistochemistry was used to detect the expression of Aurora-A in nude mice The effect of Aurora-A overexpression on the expression of VEGFR-2 and p-VEGFR-2 (Tyr1059) in KYSE150 cells was detected by Western blot. Results The total Aurora-A protein expression in Aurora-A high expression group was about 2.5-fold higher than that in negative control group and non-transfected group, suggesting that Aurora-A high expression cell line was constructed successfully. MVD and p-VEGFR-2 expression levels were significantly higher than the negative control group (P <0.01). Conclusion Aurora-A overexpression can promote angiogenesis in esophageal squamous cell carcinoma, which may be related to the activation of VEGFR-2.