甲基莲心碱对耐阿霉素人乳腺癌细胞凋亡抗性的影响

Poteintation of adriamycin-induced apoptosis by neferine in resistant human breast cancer cell line MCF-7/Adr

目的 探讨甲基莲心碱 (neferine ,Nef)对耐阿霉素人乳腺癌细胞 (MCF 7/Adr)凋亡抗性的影响及其机制。方法 应用Tunel法和PI染色流式细胞仪检测细胞凋亡 ,间接免疫荧光流式细胞仪检测P gp的表达 ,高效液相色谱法(HPLC)检测细胞内阿霉素 (ADR)的浓度。结果 ①MCF 7/Adr细胞能耐受 5mg·L- 1 ADR诱导的凋亡 ,而 1 ,5 ,1 0μmol·L- 1 Nef可使 5mg·L- 1 ADR诱导的MCF 7/Adr细胞凋亡从 7 95 %分别增加至 2 1 3 % ,2 7 9% ,61 3 % ;② 1 0μmol·L- 1 Nef可使MCF 7/Adr细胞内ADR积累由 0 52 μg·(1 0 6 cells) - 1 增加到 1 50 μg·(1 0 6 cells) - 1 ;③ 1 0 μmol·L- 1Nef作用 2 4h后 ,MCF 7/Adr细胞P gp的表达明显下降。结论 甲基莲心碱能逆转MCF 7/Adr细胞的凋亡抗性 ,其作用机制可能与抑制P gp的功能和表达、增加ADR在MCF

AIM To investigate the poteintation of adriamycin induced apoptosis by neferine in resistant human breast cancer cell line MCF 7/Adr. METHODS Apoptosis was detected by PI stain flow cytometry and Tunel assay. The intracellular adriamycin (ADR) accumulation was assayed by HPLC and the expression of P gp was determined by flow cytometry. RESULTS (1)MCF 7/Adr cells resisted the apoptosis induced by ADR while Nef augmented ADR medidated apoptosis; (2) Nef (10 μmol·L -1 ) increased the accumulation of ADR up to 2 88 fold in MCF 7/Adr cells but not in sensitive cells MCF 7/S; (3) Nef(10 μmol·L -1 ) reduced the expression of P gp in MCF 7/Adr cells. CONCLUSIONS Nef can overcome apoptosis resistance in MCF 7/Adr cells and its mechanisms are involved in the augment of ADR accumulation and the down-modulation of P gp expression in MCF 7/Adr cells.