Reversion of P-glycoprotein-mediated multidrug resistance by guggulsterone in multidrug-resistant human cancer cell lines
Reversion of P-glycoprotein-mediated multidrug resistance by guggulsterone in multidrug-resistant human cancer cell lines
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在多重耐药人癌细胞系中古古甾酮逆转 P-糖蛋白介导的多重耐药性
DOI：
10.1016/j.ejphar.2012.06.046
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发表时间：
2012-11-05
影响因子：
5
通讯作者：
Mao, Xia-Ping
中科院分区：
文献类型：
--
作者：
Xu, Hong-Bin;Xu, Lu-Zhong;Mao, Xia-Ping
多药耐药（MDR）是肿瘤化疗中的一个严重问题。本课题组前期研究表明，杜仲甾酮可通过抑制P-糖蛋白（P-gp）的功能和表达而逆转MDR。本研究旨在进一步探讨杜仲甾酮对肿瘤耐药细胞多药耐药的逆转作用。分别采用RT-PCR、pH敏感荧光探针BCECF-AM、Pgp-Glo检测系统和流式细胞术技术检测葛根素对MDR 1 mRNA表达、细胞内pH、Pgp-ATP酶活性和葡萄糖神经酰胺合成酶（GCS）表达的影响。结果表明，在2.5 ~ 80 μ M浓度范围内，guggulsterone对P-gp ATP酶活性有明显的促进作用，并呈剂量依赖性。K562/DOX细胞内pH值高于K562细胞。经guggulsterone（1、3、10、30、100 μ M）处理后，K562/DOX细胞内pH值呈剂量和时间依赖性下降。然而，目前的研究表明，guggulsterone从3到100 μ M对K562/DOX细胞的MDR 1基因表达几乎没有影响。此外，与亲本MCF-7人乳腺癌细胞相比，等基因阿霉素抗性MCF-7/DOX细胞显示GCS水平增加4.9倍。MCF-7/DOX细胞经guggulsterone（0.1、1、10 μ M）处理48 h后，GCS蛋白表达量无明显变化。银杏甾酮可能是一种有效的MDR逆转剂，其作用机制有待进一步研究。(c)2012爱思唯尔有限公司版权所有。
Multidrug resistance (MDR) presents a serious problem in cancer chemotherapy. Our previous studies have shown that guggulsterone could reverse MDR through inhibiting the function and expression of P-glycoprotein (P-gp). The present study is to further investigate the reversal effects of guggulsterone on MDR in drug-resistant cancer cell lines. The effects of guggulsterone on MDR1mRNA gene expression, intracellular pH, P-gp ATPase activity and glucosylceramide synthase (GCS) expression were assessed by RT-PCR, Laser Scanning Confocal Microscope using the pH-sensitive fluorescent probe BCECF-AM, Pgp-Glo assay system, and flow cytometric technology, respectively. The results showed that guggulsterone ranging from 2.5 to 80 mu M significantly promoted the activity of P-gp ATPase in a dose-dependent manner. The intracellular pH of K562/DOX cells was found to be higher than K562 cells. After treatment with guggulsterone (1, 3, 10, 30, 100 mu M), intracellular pH of K562/DOX cells decreased in a dose- and time-dependent manner. However, the present study revealed that guggulsterone ranging from 3 to 100 mu M had little influence on MDR1 gene expression in K562/DOX cells. Further, the isogenic doxorubicin-resistant MCF-7/DOX cells exhibited a 4.9-fold increase in GCS level as compared with parental MCF-7 human breast cancer cells. After treatment with guggulsterone (0.1, 1, 10 mu M) for 48 h, MCF-7/DOX cells were found to have no change of GCS protein expression amount. Guggulsterone might be a potent MDR reversal agent, and its mechanism on MDR needs more research. (c) 2012 Elsevier B.V. All rights reserved.