油菜素内酯对CCRF-VCR1000细胞多药抗药性的逆转作用及其机制的初步探讨  

Reversing effect of brassinolide on multidrug resistance of CCRF-VCR1000 cells and a preliminary investigation on its mechanisms

作　　者：冼励坚[1] 李永强[1] 刘然义[1] 曹弃元[1] 孙健[1] 

机构地区：[1]中山大学肿瘤防治中心,广东广州510060

出　　处：《药学学报》2005年第2期117-121,共5页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(39670839).

摘　　要：目的　了解油菜素内酯(BR)对多药抗药性细胞CCRF VCR1000耐药性的逆转及机理。方法　MTT法测定耐药因子及逆转倍数;罗丹明法测定药物积累;Sulliven方法测定拓扑异构酶II催化活性;Westernblotting方法检测p53蛋白表达。结果　CCRF VCR1000细胞对阿霉素、VP 16和VCR分别呈 153 .1, 55. 9和 8 .123 .1倍耐药。BR在不具细胞毒的浓度下 (0. 001 -10 .00μg·mL-1 )使耐药性逆转 4 .4 -11 .6倍;并使耐药细胞药物积累改善;过度表达的p53回复到敏感细胞水平。结论　在无细胞毒浓度下,BR能逆转CCRF VCR1000的耐药性;抑制P糖蛋白对药物的外排。下调异常表达的p53蛋白可能是其逆转机制之一。Aim To investigate the effect of brassinolide, a plant growth modulator, on multidrug resistance (MDR) of human T lymphoblastoid cell line CCRF-VCR 1000 which was obtained by progressively addition of vincristine (VCR) to sensitive CCRF-CEM cells, and to explore preliminarily the mechanism of reversing action. Methods MTT method was used to detect the resistant factor of resistant cell line and the reversing fold after addition of brassinolide. The intracellular accumulation of rhodamine 123, a fluorescent dye transported by P-glycoprotein was detected by flow cytometry, the catalytic activity of topoisomerase II was assessed by Sulliven method to find the effect of brassinolide on resistance. The protein expression of p53 was measured using Western blotting in the sensitive cells and resistant cells to explore the effect of brassinolide. Results The resistant factors of CCRF-VCR cells on adriamycin, VP-16 and VCR are respectively as 153.1, 55.9 and 8 123.1 folds comparing to the sensitive cell line CCRF-CEM. After treatment of brassinolide under the concentration of 0.001-10.0 μg·mL-1, the resistance of CCRF-VCR was reversed partly with the reversing folds respectively as 4.4-11.6. The intracellular accumulation of rhodamine 123 was significantly reduced in the resistant cells. After treatment of brassinolide, the accumulation increased, the level of fluorescent dye was situated between resistant cells and sensitive cells. No alteration of the catalytic activity of topoisomerase II was found among three groups. The level of protein expression of p53 in resistant cells was higher than that of sensitive cells. After brassinolide treatment, the expression of p53 in CCRF-VCR cells restored to the level of sensitive cells. Conclusion Brassinolide could effectively reverse the resistance of CCRF-VCR cells by inhibiting the effusion of drug transported by P-glucoprotein. To down regulate the abnormal expression of p53 maybe one of the mechanisms of reversing MDR for brassinolide.

关 键 词：油菜素内酯 逆转 多药抗药性 肿瘤细胞 植物生长调节剂 

分 类 号：R963[医药卫生—微生物与生化药学] R979.1[医药卫生—药理学]