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目的 :观察MCF - 7/Adr耐药细胞CD95分子的表达和功能及其与药敏细胞的差异 ,初步探讨CD95缺陷与多药耐药的潜在关系。方法 :对乳腺癌MCF - 7/Adr耐药细胞和MCF - 7药物敏感细胞进行对比研究。流式细胞仪检测CD95蛋白水平 ,RT -PCR分析CD95mRNA转录体的多态性。碘化丙啶染色和DNA琼脂糖凝胶电泳观测抗CD95单克隆抗体对MCF - 7/Adr耐药细胞的诱导凋亡效应。结果 :MCF - 7/Adr耐药细胞CD95阳性率为 5 6 6 3%明显低于MCF - 7药物敏感细胞株 (阳性率为 6 7 38% ) ;RT -PCR显示MCF - 7/Adr耐药细胞CD95mRNA同时表达两种CD95转录体 ,一种为 10 0 9bp的结构正常的分子 ,另一种为 0 7kb左右长度的可溶性CD95。MCF - 7药物敏感细胞只表达膜型CD95一种分子 ;1 0 μg/ml抗CD95McAb作用 2 4小时 ,MCF -7药物敏感细胞的凋亡率为 84 6 7% ,而MCF - 7/Adr耐药细胞的凋亡率仅为 14 37%。结论 :MCF - 7药物敏感细胞和其MCF - 7/Adr耐药亚系细胞在CD95分子多态性和介导凋亡功能上均存在显著差异 ,提示耐药细胞对CD95表达和功能的调控与药敏细胞有重要差异。
Objective: To observe the expression and function of CD95 molecule in MCF-7/Adr drug-resistant cells and the differences between them and drug sensitive cells, and to explore the potential relationship between CD95 deficiency and multidrug resistance. METHODS: A comparative study of breast cancer MCF-7/Adr-resistant cells and MCF-7 drug-sensitive cells was performed. The CD95 protein level was detected by flow cytometry, and the CD95 mRNA transcript polymorphism was analyzed by RT-PCR. Propidium iodide staining and DNA agarose gel electrophoresis were used to observe the apoptosis-inducing effect of anti-CD95 monoclonal antibody on MCF-7/Adr-resistant cells. RESULTS: The positive rate of CD95 in MCF-7/Adr resistant cells was 5666.3%, which was significantly lower than that of MCF-7 sensitive cells (positive rate was 6738%). RT-PCR showed that MCF-7/Adr resistance The cell CD95 mRNA simultaneously expresses two CD95 transcripts, one with a normal structure of 109 bp and the other with soluble CD95 with a length of about 0.7 kb. MCF-7 sensitive cells expressed only one molecule of membrane type CD95; the effect of anti-CD95 McAb at 10 μg/ml was 24 hours, and the apoptosis rate of MCF-7 sensitive cells was 84 6 7%, while MCF-7/Adr resistance The apoptotic rate of drug cells is only 14 37%. Conclusion: There are significant differences in CD95 molecular polymorphism and apoptosis-inducing function between MCF-7-sensitive cells and their MCF-7/Adr-resistant sublineage cells, suggesting that the regulation of CD95 expression and function by drug-resistant cells There are important differences in drug sensitive cells.