【作者】 孔雁；

【导师】 姜达；

【作者基本信息】 河北医科大学 ， 肿瘤学， 2006， 硕士

【摘要】 目的：通过体外实验观察沙利度胺对人乳腺癌细胞血管内皮生长因子-C(vascular endothelial growth factor-C，VEGF-C)及其受体(vascular endothelial growth factor receptor-2，VEGFR-2)的表达影响，进一步阐明沙利度胺抗血管生成作用的可能机制，探讨其用于治疗乳腺癌的可行性。 方法：选取两株经典的人乳腺癌细胞系MCF-7及MDA-MB-231，采用四甲基偶氮唑蓝(MTT)法检测沙利度胺不同浓度组(6、30、60、120μg/ml)、不同时间组(24、48、72h)对细胞存活和生长的影响情况，从而选择适当的药物浓度及作用时间。根据MTT的结果，设置对照及实验组。分别提取各组细胞总RNA，鉴定其完整性及含量，通过逆转录—聚合酶链反应(revers transcription PCR，RT-PCR)半定量检测沙利度胺处理前后两株细胞中VEGF-C、KDR mRNA的表达水平。应用免疫细胞化学(immunocytochemistry，IC)方法定性观察沙利度胺处理前后MCF-7、MDA-MB-231细胞中VEGF-C、KDR蛋白的表达情况。采用流式细胞术(flow cytometry，FCM)半定量检测沙利度胺处理前后两株细胞中VEGF-C及KDR蛋白的表达水平。 结果：1 MTT结果显示：沙利度胺在(6～120)μg/ml浓度范围内能明显抑制MCF-7和MDA-MB-231细胞体外增更多还原

【Abstract】 Objective: This experiment aimed to evaluate the effect of thalidomide on the expression of vascular endothelial growth factor-C (VEGF-C) and its receptor (vascular endothelial growth factor receptor-2, VEGFR-2) in breast cancer cell lines in vitro to further investigate possible mechanism of its role as angiogenesis inhibitor and feasibility to cure breast cancer.Methods: Selecting two-stub classic human breast cancer cell lines, MCF-7 and MDA-MB-231, using MTT assay to detect the growth rate among different thalidomide concentration groups (6、30、60、120μg/ml)and different time groups (24、48、72h), in order to choose proper drug concentration and action time. According to the result of MTT, establishing control and experimental group, extracting total RNA of each group cell, assessing the integrality and content of RNA, the level of VEGF-C、KDRmRNA expression was examined by semiquantitative RT-PCR technique in the two-stub cell lines treated before and after with thalidomide. The expression of VEGF-C、KDR protein was qualitationly studied by immunocytochemistry staining and was semi-quantitately examined by flow cytometry in cell lines treated before and after with thalidomide.更多还原