ILK在胶质瘤中表达及对VEGF、HIF1α分泌表达影响.pdfVIP

摘 要 I 摘 要 目的: 研究整合素连接激酶（ILK）在胶质瘤组织中的表达，并探讨其与胶质瘤血管生 成之间的关系 方法：1）收集不同级别人脑胶质瘤组织标本，免疫组化检测肿瘤标本 ILK、VEGF、 HIF1-α的蛋白表达并分析其相关性； 2）shRNA 转染胶质瘤细胞，并建立稳定转染的细胞系； 3）采用 RT-PCR、Western blot 检测 U251 稳定转染细胞系中 ILK 的转录及表达 情况； 4）Western blot、ELISA 检测 shRNA 敲低 ILK 后胶质瘤细胞的 VEGF、HIF-1α 的表达； 结果：ILK 在各级别胶质瘤组织中的表达有显著性差异（P0.05）,其表达量与肿瘤恶性 程度呈正相关（p＜0.05），且 VEGF、HIF1-α随着 ILK 的表达增高而增高，呈显著正相 关（r=0.975,p0.01；r=1.000,p＜0.01）；稳定转染的 U251 细胞 ILK mRNA 和蛋白表达 水平明显下降，而且 VEGF、 HIF-1α的表达量同样下降明显。 结论：ILK 与胶质瘤的血管生成过程密切相关，抑制 ILK 的表达可以有效的抑制胶质瘤 的血管生成。 关键词 ILK shRNA 免疫组化 胶质瘤 血管生成 Abstract II Abstract Objective: To study the expression of Integrin-Linked kinase (ILK) in the glioma tissues, and to explore its role in the process of glioma angiogenesis. Methods: 1) Different grades of samples of human glioma tissues were collected and immunohistochemistry staining were performed to detect the expression of ILK, VEGF and HIF1-α as well as analyze those correlations; 2) Glioma cells were transfected by shRNA and stable transfection cell lines were established; 3）The transcription and expression of ILK in the U251 stable transfection cell lines were detected by RT-PCR and western blot methods; 4) The expression of VEGF and HIF1-α in glioma cells whose ILK was knocking down by shRNA were detected through western blot methods and Elisa methods. Results: The expressions of ILK in different grades of glioma tissues had significant differences (P 0.05) and those expressions were positively correlated with the malignant degrees of the tumor (P 0.05). The protein expression levels of VEGF and HIF1-α had significant positive correlation with the expression of ILK (r = 0.975, P 0.01 and r = 1.000, P 0.01). The expression levels of mRNA and protein in stable transfection cell lines U251’s ILK decreased obviously and the expression levels of VEGF and HIF1-α also decreased significantly. Conclusion: ILK was closely related to the process of glioma angiogenesis. To inhibit the exp