环孢菌素a、雷洛昔芬及其联合应用逆转k-a细胞多药耐药的研究.docVIP

环孢菌素A、雷洛昔芬及其联合应用逆转K562/A02细胞多药耐药的研究 nbsp;nbsp;nbsp; 作者：鲍文,陈宝安,高峰,丁家华,许文林1,沈惠玲1,高冲,孙耘玉,程坚,王骏,赵刚,马燕 【摘要】nbsp; 本研究探讨环孢菌素A（CsA）、雌激素受体抑制剂雷洛昔芬及其联合应用对K562/A02细胞多药耐药的逆转作用。采用甲基四唑蓝法(MTT)测定柔红霉素(DNR)的半数抑制量，RT-PCR法检测mdr1基因mRNA表达水平，FCM检测P-gp的表达和细胞内DNR浓度。结果表明： DNR对K562/A02和K562细胞的IC50分别为23.51和0.29mg/L，雷洛昔芬(2.5 mg/L)及CsA( 1 mg/L)单用和两药联合处理K562/A02细胞时， DNR的IC50值分别为5.98、8.15和3.68 mg/L，两药对DNR作用K562细胞的IC50没有影响。CsA及雷洛昔芬单独作用后耐药株mdr1 mRNA下调微弱，联合用药下调效果明显。CsA及雷洛昔芬均可降低P-gp蛋白的表达，且具有协同作用。同时还观察到逆转剂雷洛昔芬和CsA作用后细胞内柔红霉素浓度增加，两药联合作用时效果增强。结论： CsA及雷洛昔芬均可逆转耐药,且联合作用效果增强。 【关键词】nbsp; 环孢菌素A; 雌激素受体抑制剂; 雷洛昔芬； K562/A02细胞; 多药耐药 　　Effect of Cyclosporine A, Raloxifene and Their Combination on the Reversion of Multidrug Resistace of K562/A02 Line 　　AbstractThisnbsp; study was aimed to investigate the reversible effect of cyclosporine A, raloxifene and theirnbsp;nbsp; combinationnbsp; on multidrug resistance cell line K562/A02. The IC50 (the concentration causing 50% inhibition of cell growth) of DNR were assayed by MTT method, the expression level of mdr-1 mRNA was assayed by RT-PCR,nbsp;nbsp;nbsp;nbsp; p-glycoprotein (P-gp) expressionnbsp;nbsp; and intracellular DNR concentrationnbsp;nbsp; were detected by flow cytometry. The results showed thatnbsp;nbsp;nbsp; the IC50 of DNR on K562/A02 and K562 cells were 23.51 mg/L and 0.29 mg/L,nbsp; respectively.nbsp; Thenbsp; IC50 of DNR on K562/A02 cells in treatment withnbsp; raloxifene CsAnbsp;nbsp; and both combinationnbsp; werenbsp; 5.98,8.15 and 3.68 mg/L respectively, but both drugsnbsp;nbsp; not influenced IC50nbsp; of DNR onnbsp;nbsp; K562 cells. Pretreating K562/A02 cells with raloxifene (2.5 mg/L) or CsA (1 mg/L)nbsp; for 48 hours partially restored the sensitivity of K562/A02 cells to DNR.nbsp;nbsp;nbsp; Cyclosporine A and raloxifene (alone or combination) elevated the intracellular DNR concentrationnbsp; in K562/A02,nbsp; down regulatednbsp; P-gp and mdr-1 mRNAnbsp; expressions.nbsp;nbsp; It is concluded thatnbsp; multidrug resistance (MDR) can be partially rev