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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 7/Adr细胞内的积累有关
Objective To investigate the effect and mechanism of neferine (Nef) on the apoptosis resistance of doxorubicin-resistant human breast cancer cells (MCF-7 / Adr). Methods The apoptosis was detected by Tunel method and PI staining. The expression of P gp was detected by indirect immunofluorescence flow cytometry. The concentration of adriamycin (ADR) was detected by high performance liquid chromatography (HPLC). Results ①MCF 7 / Adr cells were able to tolerate 5 mg · L-1 ADR-induced apoptosis, whereas 1, 5 and 10 μmol·L-1 Nef could induce apoptosis of MCF 7 / Adr cells induced by 5 mg · L- 795% increased to 21.3%, 27.9% and 61.3%, respectively. ② Neodymium 1 0μmol·L-1 increased ADR accumulation in MCF 7 / Adr cells from 0 52 μg · (1 0 6 cells) 1 to 1 50 μg · (1 0 6 cells) -1; ③ After the cells were treated with 1 μmol·L -1 NiF for 24 hours, the expression of P gp in MCF 7 / Adr cells was significantly decreased. Conclusion Neferine can reverse the apoptosis resistance of MCF 7 / Adr cells, and its mechanism may be related to inhibiting the function and expression of P gp and increasing the accumulation of ADR in MCF 7 / Adr cells