雷公腾内酯与多柔比星诱导耐药白血病细胞凋亡协同作用及其机制的探讨  

Exploration of the ability of triptolide to increase drug induced apoptosis in resistant leukemia cell lines and relationship with Nrf2 pathway

作　　者：陈菲莉[1] 柳约坚[1] 李蓉蔚[1] 王诗韵[1] 董慧娟[1] 周淑芸[1] 徐兵[1] 

机构地区：[1]南方医科大学南方医院血液科,广东广州510515

出　　处：《中华肿瘤防治杂志》2014年第10期728-731,共4页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(81070425);广东省科技计划(2011B031800063);南方医院院长基金(2012C007)

摘　　要：目的:以耐多柔比星(adriamycin,ADM)的人急性髓系白血病(acute myeloid leukemia,AML)耐药细胞株HL60/ADM为研究对象,探讨IC20浓度雷公腾内酯(triptolide,TPL)能否提高ADM诱导耐药白血病细胞凋亡及其与Nrf2通路的关系。方法:流式细胞仪检测空白对照组、IC20浓度TPL单药组、ADM单药组和TPL联合ADM组处理HL-60/ADM后细胞凋亡率;实时荧光定量PCR检测各个处理组作用后,Nrf2及其下游基因醌氧化还原酶(quinone oxidoreductase,NQO1)、谷胱甘肽还原酶(glutathione reductase,GSR)及血红素加氧酶1(heme oxygenase 1,HO-1)的表达水平变化;蛋白质印迹法检测各处理组作用后Nrf2蛋白表达变化。结果:TPL单药组细胞凋亡率为(5.28±0.80)%,与空白对照组的(7.09±0.46)%比较差异无统计学意义,P=0.226;但该浓度TPL可使ADM组细胞凋亡率由(19.55±1.70)%提高到(72.62±4.83)%,是ADM单药组的3.71倍,P<0.001。空白对照组、TPL单药组、ADM单药组及双药联合组Nrf2mRNA表达水平分别为1、0.742±0.052、0.619±0.042和0.241±0.010,NQO1分别为1、0.363±0.075、0.228±0.053和0.050±0.034;GSR分别为1、0.268±0.042、0.231±0.106和0.038±0.017;HO-1分别为1、0.495±0.023、0.282±0.099和0.048±0.036;各用药组Nrf2及其下游基因NQO1、GSR及HO-1的mRNA表达水平较对照组均出现显著下调,P<0.001;其中双药联合组下调程度最大。蛋白质印迹法结果显示,用药组及联合组Nrf2表达水平较对照组均有不同程度下调,其中联合组下调最为明显。结论:IC20浓度雷公腾内酯可显著提高ADM诱导耐药白血病细胞凋亡,其分子机制与下调Nrf2通路有关。OBJECTIVE：To use cell line HL60/ADM which is resistant to Adriamycin to evaluate whether low dose（IC20）of Triptolide can increase the apoptosis induced by Adriamycin and its relationship with Nrf2pathway.METHODS： HL60/ADM cells were subjected to different treatments and flow cytometry and Western blot or QT-PCR were used to determine IC20,apoptotic status and expression of Nrf2and their target genes.RESULTS：Apoptotic ratio of TPL-treated group was（5.28±0.80）%.There was no difference of apoptotic ratio between TPL-treated group and control group（7.09±0.46）%（P=0.226）but the TPL concentration could increase the apoptotic ratio of ADM group from（19.55± 1.70）%to（72.62±4.83）%（P〈0.001）.The Nrf2mRNA level of control,TPL,ADM and combination group was 1, 0.742±0.052,0.619±0.042,0.241±0.010;NQO1was 1,0.363±0.075,0.228±0.053,0.050±0.034;GSR was 1, 0.268±0.042,0.231±0.106,0.038±0.017;HO-1was 1,0.495±0.023,0.282±0.099,0.048±0.036;mRNA level of NQO1,GSR and HO-1in the treated group was significantly lower than that in control group,with the highest decline in combination group.Western blotting result showed that Nrf2of treated group was significantly lower than that of control group,with the highest decline in the combination group.CONCLUSION：Low dose（IC20）of Triptolide could increase the apoptosis induced by Adriamycin through down-regulation of Nrf2pathway.

关 键 词：雷公腾内酯 小剂量 HL-60 ADM 凋亡 Nrf2通路 

分 类 号：R733.7[医药卫生—肿瘤]