异钩藤碱对人肺腺癌细胞A549/DDP多药耐药的逆转作用

摘要

目的:探讨异钩藤碱对人肺腺癌多药耐药细胞株A549/DDP耐药的逆转作用及其可能的机制.方法:采用MTT法检测不同浓度异钩藤碱对A549细胞和A549/DDP细胞的增殖抑制效应;MTT法检测顺铂(cisplatin,DDP)、长春新碱(vincristine,VCR)、依托泊苷(etoposide,VP-16)对A549,A549/DDP细胞增殖的抑制作用及加用异钩藤碱时的变化;流式细胞仪检测钩藤碱对VCR引起细胞凋亡的影响,采用电感耦合等离子质谱法检测细胞内顺铂蓄积浓度.结果:1.5 mg·L-1及更低浓度的异钩藤碱对A549和A549/DDP细胞无明显细胞毒性作用.A549/DDP对VCR,VP-16,ADM和DDP的耐药倍数分别为19.58,11.97,21.74和24.29.加入0.5,1.0和1.5 mg·L-1异钩藤碱后,A549/DDP对VCR,VP-16和DDP的耐药逆转倍数均成倍增加.加入1.5 mg·L-1异钩藤碱后,A549/DDP对上述化疗药物的凋亡率提高更明显.1.5 mg·L-1异钩藤碱可明显提高DDP在A549/DDP细胞内的蓄积.结论:异钩藤碱可以逆转A549/DDP细胞的多药耐药,逆转效果与药物浓度有关,逆转机制可能与增加细胞内化疗药物蓄积和增强化疗药物诱导的细胞凋亡有关.%Objective: To investigate the reversal effect of isorhynchophylline on multidrug resistance of hu-man adenocareinoma of lung cell line A549/DDP. Methods:The effects of isorhynchophylline on the proliferation of A549 and A549/DDP cells were observed by MTT assay. The inhibitory effects of cisplatin (DDP), vincristine (VCR), adriamycin (ADM) and etoposide (VP-16) used alone or in combination with isorhynchophylline on the proliferation of A549 and A549/DDP cells were evaluated by MTT assay. VCR induced apoptosis were determined by flow eytometry (FCM). The effects of isorhynchophylline on DDP accumulation in A549/DDP cells was deter-cytotoxieity to A549 and A549/DDP cells. The resistances of A549/DDP cells to VCR, VP-16, ADM and DDP were 19.58,11.97,21.74 and 24.29 times as those of A549 cells. After 0.5 mg. L-t isorhynchophylline was add-ed, the chemo-sensitivity of A549/DDP cells to VCR, VP-16 and DDP was increased as 1.27,1.81 and 2.19 in A549/DDP cells by 1.79, 2.46 and 9.30 folds as those of control group. Conclusion: Isorhynchophylline could moderately reverse the multidrug resistance of A549/DDP cells through increasing the accumulation of chemo-drugs in cells and promoting apoptosis.