【摘要】 目的探讨不同浓度顺铂作用不同时间诱导敏感和耐药卵巢癌细胞系发生凋亡变化规律和凋亡途径相关蛋白的表达。方法选择卵巢癌顺铂耐药细胞系A2780/DDP和敏感细胞系A2780为研究对象。在细胞培养基础上,运用MTT比色法、原位标记法、常规免疫组化和蛋白印迹等方法,观察不同剂量的顺铂(5、10、20、40μmol/L)和作用不同时间(24、48、72h)对A2780和A2780/DDP细胞凋亡的影响和此动态变化过程中半胱天冬氨酸蛋白酶2(Caspase2)和3(Caspase3)表达。结果顺铂对A2780和A2780/DDP细胞系凋亡的诱导呈时间和剂量依从性。细胞凋亡蛋白Caspase2和Caspase3的活化促进顺铂诱导的凋亡,其表达呈现出对顺铂作用时间和剂量的依从性。结论(1)在顺铂耐药机理中,药物在体内分布和代谢,对血药有效浓度的影响和细胞对药物排斥引起顺铂进入细胞存在一定的阻碍可能是降低顺铂敏感性的因素。(2)Caspase2和caspase3蛋白活化延后和活性降低可能是影响耐药A2780/DDP细胞凋亡的原因。更多还原

【Abstract】 Objective To investigate the law during cisplatin-induced apoptosis in cisplatin-resistant and sensitive human ovarian cancer cell line A2780 and A2780/DDP and correlative protein expressions in mitochondria apoptotic pathway.Methods Two ovarian cancer cell lines,one is cisplatin-sensitive line A2780 and the other is resistant-line A2780/DDP,were selected.Based on cell culture,the influence of varied doses(5,10,20,40μM) and time(24,48,72h) of cisplatin on apoptosis in A2780 and A2780/DDP cell lines and protein expressions of Caspase-2(cystein-containing,aspartat-specific proteases) and Caspase-3 were detected by using MTT,in situ cell apoptosis detection,immunohistochemistry technology and western blotting.Results Cisplatin-induced apoptosis in A2780 and A2780/DDP cell line depended on the time and doses of used cisplatin.Activated Caspase-2 and Caspase-3 proteins promoted apoptosis and Caspase-2 and Caspase-3 protein expressions also depended on the time and doses of used cisplatin.Conclusion (1) In pathogenesis of cisplatin resistance,the pharmaceutical distribution and metabolize and the obstacle of entering cells for rejection in vivo,may lead to decreased cisplatin-sensitivity.(2) The delayed and decreased activation of Caspase-2 and Caspase-3 proteins may have negative influence on apoptosis in resistant A2780/DDP cell.更多还原