摘要
[目的]探讨mdr1基因编码的P-糖蛋白(mdr1/P-gp)在阿霉素(ADR)诱导的卵巢癌多药耐药细胞中的作用。[方法]MTT法检测药物对敏感细胞株(SK-OV-3)和诱导的耐药细胞株(SK-OV-3/adr)的半数致死浓度(IC50),计算耐药指数(RI)和加逆转剂后的逆转倍数,荧光显微镜观察不同时间细胞内的药物含量变化,荧光分光光度法测定细胞内ADR的含量,RT-PCR和Western blot法检测细胞耐药前后mdr1/P-gp的变化。[结果]对阿霉素、长春新碱、紫杉醇和足叶乙甙,SK-OV-3/adr细胞株的IC50均有明显增加,RI分别为58.4、122.6、41.0和43.7,逆转倍数分别为20.5、66.9、11.3和18.1。30、60、120 min SK-OV-3细胞株ADR含量分别是SK-OV-3/adr细胞株的3.7[(191.9±1.7)/(51.7±6.3)ng/mL]、5.9[(285.3±4.0)/(48.2±2.8)ng/mL]和8.6[(447.8±1.3)/(52.0±1.8)ng/mL]倍,二者比较差异有非常显著性意义(P<0.01)。P-gp竞争性逆转剂(Ver)作用30、60、120 min后SK-OV-3/adr细胞ADR含量提高了1.6、2.7和5.1倍,与逆转前相比差异有非常显著性意义(P<0.01)。RT-PCR和Western blot结果显示SK-OV-3/adr细胞mdr1/P-gp均表达阳性,SK-OV-3细胞mdr1/P-gp表达阴性。[结论]mdr1/P-gp在ADR诱导的SK-OV-3/adr细胞多药耐药机制中起重要作用。
[ Objective ] To investigate the effect of P- glucoprotein （mdrl/P -gp）encoded by mdrl gene on drug resistant cells of ovarian cancer induced by Adriamycin（ADR）. [ Methods] MTT assay was employed to test the lethal concentration 50（ IC50 ） of sensitive germline （ SK - OV - 3） and ovarian cancer cell subline induced by ADR （ SK - OV - 3/adr） treated by 4 drugs,resistance index and reverse multiple were calculated. The changes of drug content were observed by fluores- cence microscope,the intracellular accumulation of ADR was evaluated by fluorospectrophotometry,the changes of mdr 1/P - gp resistance were observed respectively by RT - PCR and Western blot. [ Results ] After treatment of Adriamycin, Vincristine,Paclitaxel and Etoposide, the IC50 of SK - OV - 3/adr increased significantly, the resistance indexes （RIs） were 58.4,122.6,41.0 and 43.7, respectively. ADR contents of SK -OV -3 detected by fluorospectrophotometry in 30min, 60rain and 120 min were 3.7,5.9,8.6 times, respectively （ SK - OV - 3 vs SK - OV - 3/adr,P 〈 0.01 ）. After treated by P - gp competively reverse agent（Ver） in 30min ,60 rain and 120 rain ADR contents of SK - OV - 3/adr were 1.6,2.7 and 5.1 times higher （P 〈0.01 ）. The mdr 1/P -gp expression of SK -OV -3/adr cell was observed by RT -PCR and West-ern blot,but lost in SK -OV -3 cell. [ Conclusion] mdr1/P -gp plays a role in the muhidrug resistance of SK -OV -3/ adr cell induced by ADR.
出处
《大连医科大学学报》
CAS
2010年第1期35-39,44,共6页
Journal of Dalian Medical University
