帕博西尼逆转人乳腺癌MCF-7/DOX细胞对多柔比星的耐药性  

Reversal effect of palbociclib on the resistance of breast cancer MCF-7 cells to doxorubicin

作　　者：赵国智[1] 温纯洁[1] 邹镇[1] 吴兰香[1] 周宏灏[1,2] 

机构地区：[1]重庆医科大学生命科学研究院,重庆400016 [2]中南大学湘雅医学院临床药理研究所,湖南长沙410078

出　　处：《肿瘤》2017年第8期831-839,共9页Tumor

基　　金：国家自然科学基金资助项目(编号:81473284)~~

摘　　要：目的:研究细胞周期蛋白依赖性激酶4/6(cyclin-dependent protein kinase4/6,CDK4/6)抑制剂帕博西尼(palbociclib,PAL)对乳腺癌MCF-7/多柔比星(doxorubicin,DOX)细胞耐药性的逆转作用,并探讨药物转运体ATP结合盒蛋白B1(ATP binding cassette protein B1,ABCB1)在其中的作用机制。方法:采用CCK-8法检测PAL对亲本MCF-7及耐药MCF-7/DOX细胞DOX敏感性的影响;FCM法检测PAL对MCF-7及MCF-7/DOX细胞中DOX蓄积浓度的影响。构建稳定转入ABCB 1基因的MDCKⅡ细胞模型,采用药物跨膜转运实验检测PAL对药物转运体ABCB1介导的DOX外排功能的影响。实时荧光定量PCR和蛋白质印迹法检测PAL对MCF-7及MCF-7/DOX细胞中ABCB1 mRNA及蛋白表达水平的影响。结果:0.2与0.4μmol/L的PAL可使DOX对MCF-7/DOX细胞的半数抑制浓度(half maximal inhibitory concentration,IC_(50))分别降低39%和51%(P值均<0.05),但对MCF-7细胞的IC_(50)值无明显影响;0.4μmol/L PAL可使MCF-7/DOX细胞内DOX的蓄积浓度上调82%(P<0.05),但对MCF-7细胞中DOX的蓄积浓度无明显影响。PAL对ABCB1介导的DOX外排功能有明显抑制作用(P<0.05),但对MCF-7及MCF-7/DOX细胞中ABCB1 mRNA与蛋白的表达水平均没有明显的调节作用。结论:PAL可以通过抑制ABCB1转运体的功能而逆转乳腺癌MCF-7细胞DOX耐药性,提示PAL与DOX联用有望提高临床乳腺癌DOX的治疗效果,优化临床治疗方案。Objective： To study the reversal effect of cyclin-dependent protein kinase 4/6 （CDK4/6） inhibitor palbociclib （PAL） on the resistance Of breast cancer MCF-7/doxorubicin （DOX） cells to DOX, and to explore its mechanism from the view of drug efflux transporter ATP binding cassette protein B1 （ABCB1）.Methods： The effect of PAL on the sensitivity of parental MCF-7 and resistant MCF-7/DOX cells to DOX was detected by CCK-8 method. The effect of PAL on intracellular accumulation of DOX in MCF-7 and MCF-7/DOX cells was determined by FCM analysis. The MDCK II cell model stably transfected with ABCB1 gene was constructed, then the effect of PAL on the ABCBl-mediated efflux function of DOX was detected by drug transmembrane bi-direction transport assay. The effect of PAL on the expressions of ABCB1 mRNA and protein in MCF-7 and MCF-7/DOX cells was detected by real-time fluorescent quantitative PCR and Western blotting, respectively.Results： After treatment with 0.2 and 0.4 μmol/L PAL, the half maximal inhibitory concentration （IC50） values of DOX in MCF-7/DOX cells were decreased by 39% and 51%, respectively （both P 〈 0.05）, but the IC50 value of DOX in MCF-7 cells had no obvious change. In MCF-7/DOX cells treated with 0.4 μmol/L PAL, the intracellular accumulation of DOX was increased by 82% （P 〈 0.05）, but the concentration of DOX in MCF-7 cells did not change. PAL significantly inhibited the ABCB1 -mediated efflux of DOX, but had no obvious regulatory effect on the expressions ofABCB1 mRNAand protein in MCF-7 cells and MCF-7/DOXcells. Conclusion： PAL can reverse DOX-resistance of breast cancer cells by inhibiting the function of drug transporter ABCB1, which suggests that coadministration of PAL and DOX maybe improve the efficacy of DOX in breast cancer and optimize the clinical treatment.

关 键 词：乳腺肿瘤 帕博西尼 多柔比星 耐药性 ABCB1 

分 类 号：R737.9[医药卫生—肿瘤]