摘要
目的探讨PI3K抑制剂对三阴性人乳腺癌细胞多药耐药的逆转作用及其与PI3K/Akt信号转导关系。方法采用MTT法测定三阴性乳腺癌细胞系MBA-MB-231及其多药耐药细胞株MBA-MB-231/ADR对经典化疗药物阿霉素的半数抑制浓度(IC50),以及MBA-MB-231/ADR细胞株在PI3K抑制剂LY294002给药前后对阿霉素的敏感性差异。Western Blot检测MDA-MB-231/ADR细胞株LY294002给药前后,P-Akt、Akt、IκBα、NF-κB、P-gp和cleaved-caspase-3表达水平差异。结果 1MBA-MB-231和耐药细胞株MBA-MB-231/ADR对阿霉素的IC50分别为(0.36±0.03)μmol/L和(16.77±1.57)μmol/L,耐药倍数为46.58倍。2LY294002给药后,耐药细胞株MBAMB-231/ADR对阿霉素的IC50降至(2.59±0.07)μmol/L,耐药倍数降至6.475,对药物敏感性显著增加。3耐药细胞株MBA-MB-231/ADR中PI3K/Akt和NF-κB通路被激活,细胞耐药相关蛋白P-gp处于高表达水平;LY294002处理之后,MBA-MB-231/ADR细胞中PI3K/Akt和NF-κB通路被抑制,P-gp表达水平下调,细胞凋亡相关的cleaved-caspase-3表达上调。结论 PI3K抑制剂LY294002能有效逆转耐药细胞株MBA-MB-231/ADR的耐药性,从而增强耐药细胞MBA-MB-231/ADR对化疗药物阿霉素的敏感性。该研究结果提示LY294002通过阻断PI3K/Akt通路抑制下游NF-κB通路的活化和P-gp表达上调,从而参与MBA-MB-231/ADR耐药性的逆转。
Objective The aim of this study is to investigate the reversal effect of multidrug resistance and its signal transduction mecha-nism of human breast cancer by inhibiting the PI3K/ Akt signaling pathway with PI3K Inhibitor. Methods Cells were treated with or without LY294002（the PI3K inhibitor）. MTT assay was used to determine the half inhibitory concentration（IC50）of breast cell lines MBA - MB - 231 and MBA - MB - 231 / ADR to Adriamycin. Western Blot was applied to analyze the expression levels of P - Akt,Akt,IκBα,NF - κB,P - gp and caspase - 3 in MDA - MB - 231 / ADR cells. Results ①The IC50 of MBA - MB - 231and MBA - MB - 231 / ADR cells to Adriamycin were （0. 36 ± 0. 03）μmol/ L and（16. 77 ± 1. 57）μmol/ L. The reversal fold of MCF - 7 / ADR was 46. 58. ②The IC50 of MBA - MB - 231 / ADR reduced to（2. 59 ± 0. 07）μmol/ L and the reversal fold dropped to 6. 475 after treating with LY294002. ③Activation of the PI3K/ Akt signal path-way was observed in MBA - MB - 231 / ADR cells,which showed up - regulation of p - Akt,P - gp and NF - κB expression levels. However,af-ter treatment of LY29400,the PI3K/ Akt pathway inhibitor,P - gp and NF - κB were down - regulated,which suggested the PI3K/ Akt pathway was inhibited. Conclusion LY294002 could effectively reverse the multidrug resistance of MBA - MB - 231 / ADR cells by inhibiting the PI3K/Akt signaling pathway.
出处
《临床和实验医学杂志》
2015年第1期1-4,共4页
Journal of Clinical and Experimental Medicine
基金
2012年广东省医学科研课题(项目编号B2012392)
关键词
乳腺癌
PI3K/AKT通路
多药耐药
逆转
Breast cancer
PI3K Inhibitor
PI3K/Akt pathway
Multidrug resistance
Reversal effect
