摘要
目的 研究不同诱导方式建立的卵巢癌顺铂耐药细胞株的耐药机理。方法 采用大剂量冲击法和浓度梯度递增法建立卵巢癌顺铂耐药细胞株Skov3/CDDP P和Skov3/CDDP 5 0。结果Skov3/CDDP P和Skov3/CDDP 5 0的耐药指数分别为 3.7和 48.6 ,伴有形态改变、生长减慢、细胞周期改变等。耐药细胞株的细胞内药物浓度降低 ,与多药耐药相关基因 (MDR1)、多药耐药相关蛋白(MRP)和肺耐药蛋白 (LRP)的表达增强有关。谷胱甘肽S转移酶 (GST π)的表达无明显变化 ,拓朴异构酶Ⅱ (TOPOⅡ )活性轻度下降。不同诱导方式存在差异。结论 顺铂耐药涉及多个相关基因的表达 ,持续诱导方式容易产生耐药。
Objective To investigate the mechanism responsible for acquired cisplatin resistance induced in human ovarian cancer cell lines by different methods. Methods Two resistant cell lines, Skov3/CDDP P and Skov3/CDDP 50, were established by pulse or stepwise exposures of ovarian cancer cell line Skov3 to cisplatin (CDDP) for 10 12 months with the drug sensitivity monitored by MTT test. The growth rate and cell cycle were compared. The intracellular drug concentration was measured by FACS after 1 hour incubation in 20 μmol/L ADM. The drug resistance associated genes: MDR1, MRP, LRP and GST π were monitored by RT PCR and western blotting. Results The resistance indexes (RI) of Skov3/CDDP P and Skov3/CDDP 50 were 3.7 and 48.6 to cisplatin, 4.0 and 33.0 to Taxol, 2.2 and 7.3 to adriamycin, 1.5 and 3.4 to VP16, and the intracellular ADM concentration was lowered by 34.6% and 47.2% respectively. Both resistant cell lines grew slowly and exhibited changes in morphology and cell cycle. The expression of the resistance associated genes MDR1, MRP and LRP was enhanced in both resistant cell lines. However, Skov3/CDDP 50 showed greater increase in MDR1 but Skov3/CDDP P showed more in MRP. There were no significant changes in GST π expression either at mRNA or protein level. The TOPO Ⅱ activity decreased slightly in both resistant cell lines. Conclusion Stepwise induction of cisplatin resistance in ovarian cancer is more readily acquired in which multiple drug resistance associated genes are involved.
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2001年第4期305-308,共4页
Chinese Journal of Oncology
基金
全军“九五”医学科研规划重点课题资助项目(96Z055)
关键词
卵巢肿瘤
顺铂
多药耐药
基因表达
Ovarian neoplasms
Cisplatin
Multidrug resistance
Gene expression
