化疗药物对结肠癌细胞SW480中免疫共刺激分子B7-H1的诱导及机理

发布时间：2018-02-04 03:45

本文关键词： 结肠癌 SW480 B7-H1　出处：《华中科技大学》2010年硕士论文　论文类型：学位论文

【摘要】：目的:研究常用化疗药物5-氟尿嘧啶和紫杉醇在结肠癌细胞中对免疫共刺激分子B7-H1表达的诱导及机理。方法:5-氟尿嘧啶和紫杉醇处理SW480细胞24小时后用RT-PCR检测B7-H1的mRNA,48小时后用免疫荧光和流式细胞仪检测B7-H1的蛋白。结果:5-氟尿嘧啶和紫杉醇在SW480细胞中可引起免疫共刺激分子B7-H1的表达:用5-氟尿嘧啶( 10μg/ml )和紫杉醇( 2.5μM )处理结肠癌SW480细胞48小时,可用免疫荧光定性检测到B7-H1蛋白的免疫荧光信号,也可以用流式细胞术半定量检测到增加的B7-H1蛋白。然而,半定量RT-PCR检测发现,B7-H1的mRNA在5-氟尿嘧啶( 10μg/ml )和紫杉醇( 2.5μM )刺激SW480细胞24小时后,并没有像蛋白一样出现增加。在本实验作为引起B7-H1蛋白增加阳性对照的γ-干扰素也不引起B7-H1 mRNA的增加。结论:化疗药物在治疗肿瘤的同时会引起B7-H1蛋白的表达,这可能与化疗不确切的治疗效果相关,同时我们也证实B7-H1分子的转录后调控在蛋白表达中起重要作用。
[Abstract]:Aim: to investigate the induction and mechanism of immunostimulatory molecule B7-H1 by 5-fluorouracil and paclitaxel in colon cancer cells. Methods SW480 cells were treated with 10% 5-fluorouracil and paclitaxel for 24 hours. The mRNA of B7-H1 was detected by RT-PCR. After 48 hours, the protein of B 7 H 1 was detected by immunofluorescence and flow cytometry. Results the expression of B7-H1 was induced in SW480 cells by using 5 fluorouracil (10 渭 g / ml) and paclitaxel (10 渭 g / ml). SW480 cells were treated with 2.5 渭 M for 48 hours. The immunofluorescence signal of B7-H1 protein could be detected qualitatively by immunofluorescence, and the increased B7-H1 protein could also be detected by flow cytometry. However, semi-quantitative RT-PCR was used to detect B7-H1 protein. B7-H1 mRNA was stimulated by 5-fluorouracil (10 渭 g / ml) and paclitaxel (2.5 渭 M) for 24 hours. There was no increase in B7-H1 protein, nor did interferon 纬 -interferon (IFN- 纬), which was used as a positive control to induce B7-H1 protein increase, increase B7-H1 mRNA in this experiment. Conclusion: chemotherapy drugs can induce the expression of B7-H1 protein at the same time, which may be related to the inexact therapeutic effect of chemotherapy. We also confirm that the posttranscriptional regulation of B _ 7-H _ 1 plays an important role in protein expression.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R392

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