三磷酸肌醇与川芎嗪通过钙通道影响肝癌耐药细胞内药物蓄积

摘要

目的 探讨三磷酸肌醇与川芎嗪对人肝癌多药耐药细胞BEL-7402/ADM中多柔比星蓄积的影响,推测钙通道参与的机制.方法 将人肝癌多药耐药细胞BEL-7402/ADM分成4组:对照组、维拉帕米组、川芎嗪组和三磷酸肌醇组,噻唑蓝(MTT)法测定4组细胞经不同浓度多柔比星作用后增殖被抑制情况,计算半数抑制浓度(IC50).荧光显微镜观察4组细胞多柔比星蓄积程度.结果 ①川芎嗪组多柔比星的IC50值为对照组的(65.35±0.24)％(P＜0.05),维拉帕米组为对照组的(64.89±0.18)％ (P＜0.05),三磷酸肌醇组为对照组的(337.60±0.12)％ (P＜0.05)；②川芎嗪组、维拉帕米组荧光强度明显高于对照组,三磷酸肌醇组荧光强度明显低于时照组.结论 三磷酸肌醇可降低人肝癌多药耐药细胞内多柔比星蓄积,增加多柔比星对人肝癌多药耐药细胞BEL-7402/ADM的IC50值,川芎嗪具有相反作用.推测三磷酸肌醇可能通过诱导人肝癌多药耐药细胞内钙激活P糖蛋白多药耐药,川芎嗪可逆转该过程.%Objective To investigate the effects of inositol triphosphate and tetramethyipyrazine on the accumulation of adriamycin in BEL-7402/ADM cells and the potential role of calcium involved. Methods BEL-74O2/ADM cells were divided into four groups: blank group,inositol triphosphate(IP3) group,tetramethylpyrazine(TMP) group and verapamil( VKP) group. MTT was used to detect inhibition of cell proliferation, and 50% inhibiting concentration value (IC50) was i-alculated. The intensity of adriamycin accumulation in the cells was observed by fluorescence microscope. Results ①IC50 value of the ceils treated with TMP.VRP and IP3 was(6S. 35±0. 24)% , (64. 89±0. 18)% and(337.60±0. 12)% (all P<0.05) of that in the blank group, respectively. ②The fluorescence intensity of the cells treated with TMP or VRP was significantly higher than the blank group,while that of the cells treated with IP3 was significantly lower than the blank group. Conclusion 1P3 reduces the accumulation of adriamycin in BEL-7402/ADM cells and increases the IC50 of adriamycin,while TMP presents the opposite effect,which suggesting that IP3 could activate P-gp by increasing intracellular calcium, but TMP reverses this process.q