甲孕酮联合苦参碱对耐药细胞株K562/AO2多药耐药逆转作用的研究

目的应用甲孕酮(MPA)联合苦参碱(MAT)逆转人白血病细胞株K562/AO2对阿霉素的耐药性,观察联合用药逆转耐药的疗效。方法用MTT法检测MPA与MAT的细胞毒性,荧光分光光度法检测各组细胞内药物(ADM)浓度的改变,流式细胞仪测定细胞凋亡的情况,流式细胞术检测细胞的p-gp表达情况。结果(1)确定了甲孕酮及苦参碱对K562/AO2细胞的非细胞毒性剂量和低细胞毒性剂量,确定了非细胞毒性剂量的两药联合应用后未出现毒性叠加。(2)在与ADM合用时,K562/AO2+甲孕酮及K562/AO2+苦参碱组,细胞凋亡百分率均高于K562/AO2耐药组(P<0.05,P<0.01),但均低于K562/AO2+甲孕酮+苦参碱组(P<0.01,P<0.05)。(3)与K562比较,K562/AO2细胞中P-gP呈高表达;与K562/AO2耐药组比较,K562/AO2十苦参碱组及K562/AO2+甲孕酮组P-gp表达量降低(P<0.01),但当两者联合应用时作用大于两者单独作用。(4)与ADM组比较苦参碱、甲孕酮均能提高K562/AO2细胞内ADM浓度,P均<0.01,两者联合应用时作用大于两者单独作用。结论非细胞毒性剂量的...

Experimental study of reversal effect of multidrug resistance by medroxyprogesterone acetate and matrine on K562/AO2 cell line

Objective: To observe the reversal effect of combination of medroxyprogesterone acetate and matrine on K562/AO2 cells.Methods: Cell toxicity of medroxyprogesterone acetate and matrine on tumor cells were determined by MTT assay.Intracellular drug concentration and cell apoptosis percentage were determined by spectrophotofluorometry and flow cytometry respectively.Changes of p-gp were determined by flow cytometry.Results:(1) Non-cytotoxic dosage and low-cytotoxic dosage of medroxyprogesterone acetate and matrine were determined by MTT assay.There was no addition toxic when Non-cytotoxic of medroxyprogesterone acetate was combined with matrine.(2) Used along with ADM,cell apoptosis percentage of K562/AO2 with medroxyprogesterone acetate or matrine was higher than that of K562/AO2(P<0.05,P<0.01respectivly),but still lower than that of K562/AO2 with medroxyprogesterone acetate and matrine(P<0.01,P<0.05respectively).(3) Compared with K562 cells,p-gp overexpress in K562/AO2;intracellular p-gp content were lower in K562/AO2 with medroxyprogesterone acetate and K562/AO2 with matrine(P<0.01),but the p-gp expressions in K562/AO2 with medroxyprogesterone acetate and matrine were lower than those of K562/AO2 with medroxyprogesterone acetate or matrine respectively.(4) Compared with ADM,medroxyprogesterone acetate and matrine can significantly enhance intracellular ADM concentration(P<0.01respectively).The effect of combination of two agents was superior to that of single uses.Conclusion: Non-cytotoxic dosage of medroxyprogesterone acetate and matrine can partly reverse the MDR of K562/AO2.The effect of combination of two agents was superior to that of single uses due to their coordinate effect;and consequently,the adriamycin related cytotoxicity that shows in cellular apoptosis ratio in K562/AO2 is harply ascended.Overexpression of p-gp in K562/AO2 is related to mechanism of MDR.