甲基莲心碱对乳腺癌MCF-7/Adr 细胞MDR逆转的研究

 目的 探讨甲基莲心碱（Neferine，Nef）对耐药人乳腺癌细胞增殖，细胞内ADM积聚浓度及mdr-1／P-gp表达的影响。方法 采用MTT法测定细胞毒作用，高效液相色谱法测定细胞内ADM积聚浓度，RT—PCR技术及蛋白质印迹技术检测mdr-1／P-gp表达。结果 10μg／ml Nef＋ADM组细胞的抑制率及细胞内ADM积聚浓度比ADM组高（P〈0.01）；20μg／ml Nef＋ADM组细胞抑制率及细胞内ADM积聚浓度较5μg／ml异博定＋ADM组及10μg／ml Nef＋ADM组均高（P〈0．01）。10μg／ml Nef＋ADM组MCF-7／Adr细胞mdr-1 mRNA及P-gp表达比ADM组明显下降（P〈0．01）。20μg／ml Nef＋ADM组细胞mdr-1mRNA及P-gp表达较10μg／ml Nef＋ADM组明显下调（P〈0．01）。结论 Nef能抑制耐药人乳腺癌细胞增殖。Nef能增加MCF-7／Adr细胞内ADM积聚浓度。Nef通过降低耐药人乳腺癌细胞mdr-1 mRNA及P-gp的表达逆转MDR。

The Study of Neferine on Multidrug Resistance Reversing of the Human Breast Cancer Cell Line MCF-7/Adr

Objective 　To investigate the effect of neferine (Nef ) on the proliferation , intracellular ADM concent ration and the expression of mdr-1/P-gp in human breast cancer cell line MCF-7/ Adr. Methods 　MTT method was used to observe cell proliferation. High performance liquid chromatography was used to detect int racellular ADM concent ration. RT-PCR and Western-blotting were used to detect mdr-1/P-gp expression. Results 　The inhibiting rate and int racellular ADM concent ration of 10μg/ ml Nef adding ADM group were significantly higher than those of ADM group ( P < 0. 01) . The inhibiting rate and intracellular ADM concent ration of 20μg/ ml Nef adding ADM group were significantly higher than those of 5μg/ ml verapamil adding ADM group and 10μg/ ml Nef adding ADM group ( P < 0. 01) . The mdr-1 mRNA and P-gp expression of 10μg/ml Nef adding ADM group was significantly lower than those of 60μg/ml ADM group ( P < 0. 01) . The mdr-1 mRNA and P-gp expression of 20μg/ml Nef adding ADM was significantly lower than that of 10μg/ml Nef adding ADM group group ( P < 0. 01) . Conclusion 　Neferine can inhibit the proliferation of human breast cancer cell line MCF-7/Adr. Neferine can improve the int racellular ADM concent ration. Neferine can reduce mdr-1 mRNA and P-gp expression and consequently reverse MDR.