摘要
目的通过不同诱导方式建立紫杉醇耐药的卵巢癌细胞系,探讨耐药机制。方法分别采用紫杉醇大剂量诱导法和小剂量间歇诱导法诱导卵巢癌细胞系SKOV3,建立卵巢癌紫杉醇耐药细胞系SKOV3/Taxol-P和SKOV3/Taxol-25。倒置显微镜进行形态学观察;细胞计数观测生长增殖规律;MTT法检测IC50和RI;流式细胞仪检测细胞周期分布;RT-PCR法检测耐药相关基因MDR1、MRP1、LRP1、GST-π等mRNA的表达。结果SKOV3/Taxol-P和SKOV3/Taxol-25的耐药指数分别为261.98±32.89和622.76±71.37,均伴有细胞形态、生长增殖、细胞周期的改变。SKOV3/Taxol-25的MDR1、LRP1表达增强,MRP1、GST-π表达均下调;SKOV3/Taxol-P中4种基因表达无改变。结论不同方式诱导的细胞系存在着差异。紫杉醇耐药涉及多个耐药基因和因素的变化。间歇诱导方式更易产生耐药。
Objective To explore the different mechanism of resistance to Taxol in human epithelial ovarian cancer cell lines by two different methods. Methods Two resistant cell lines were established： SKOV3/Taxol-P and SKOV3/Taxol- 25. They were induced from epithelial ovarian cancer cell line SKOV3 respectively by pulse and intermittent exposures to Taxol for 16 months. The biological characteristics of all the cell lines were determined by microscopy and cell counting. Drug sensitivity was monitored by MTT assay. The cell cycle was detected by flow cytometry. The mRNA expression of MDRI , MRPI , LRPI and CST-π was determined by RT-PCR. Results The resistance indexes of SKOV3/Taxol-P and SKOV3/Taxol-25 were 261.98 ± 32.89 and 622.76 ± 71.37, respectively. Both resistant cell lines had some changes in biological characteristics in which SKOV3/Taxol-25 showed more significant changes. The expression of MDRI and LRPI was up-regulated, and the expression of MRPI and C, ST-π was down-regulated in SKOV3/Taxol-25. However, SKOV3/Taxol-P showed no significant changes in MDRI , MRPI , LRPI and C, ST-π. Conclusion There are many differences between the two resistant cell lines that involved in multi-gene changes. The intermittent administration hasgreater trend to develop resistance.
出处
《基础医学与临床》
CSCD
北大核心
2006年第3期284-289,共6页
Basic and Clinical Medicine
基金
北京协和医院重点基金(200203*)
