黄芪多糖对体外 H22 肝癌细胞 P-糖蛋白外排泵功能及蛋白表达的影响。
Effects of Astragalus polysaccharides on P-glycoprotein efflux pump function and protein expression in H22 hepatoma cells in vitro.
机构信息
Clinical Pharmacy and Pharmacology Research Institute, The Second Xiangya Hospital, Central South University, Changsha, Hunan, China.
出版信息
BMC Complement Altern Med. 2012 Jul 11;12:94. doi: 10.1186/1472-6882-12-94.
BACKGROUND
Astragalus polysaccharides (APS) are active constituents of Astragalus membranaceus. They have been widely studied, especially with respect to their immunopotentiating properties, their ability to counteract the side effects of chemotherapeutic drugs, and their anticancer properties. However, the mechanism by which APS inhibit cancer and the issue of whether that mechanism involves the reversal of multidrug resistance (MDR) is not completely clear. The present paper describes an investigation of the effects of APS on P-glycoprotein function and expression in H22 hepatoma cell lines resistant to Adriamycin (H22/ADM).
METHODS
H22/ADM cell lines were treated with different concentrations of APS and/or the most common chemotherapy drugs, such as Cyclophosphamid, Adriamycin, 5-Fluorouracil, Cisplatin, Etoposide, and Vincristine. Chemotherapeutic drug sensitivity, P-glycoprotein function and expression, and MDR1 mRNA expression were detected using MTT assay, flow cytometry, Western blotting, and quantitative RT-PCR.
RESULTS
When used alone, APS had no anti-tumor activity in H22/ADM cells in vitro. However, it can increase the cytotoxicity of certain chemotherapy drugs, such as Cyclophosphamid, Adriamycin, 5-Fluorouracil, Cisplatin, Etoposide, and Vincristine, in H22/ADM cells. It acts in a dose-dependent manner. Compared to a blank control group, APS increased intracellular Rhodamine-123 retention and decreased P-glycoprotein efflux function in a dose-dependent manner. These factors were assessed 24 h, 48 h, and 72 h after administration. APS down regulated P-glycoprotein and MDR1 mRNA expression in a concentration-dependent manner within a final range of 0.8-500 mg/L and in a time-dependent manner from 24-72 h.
CONCLUSION
APS can enhance the chemosensitivity of H22/ADM cells. This may involve the downregulation of MDR1 mRNA expression, inhibition of P-GP efflux pump function, or both, which would decrease the expression of the MDR1 protein.
背景
黄芪多糖（APS）是黄芪的活性成分。它们已经被广泛研究，特别是在免疫增强特性、对抗化疗药物副作用的能力以及抗癌特性方面。然而，APS 抑制癌症的机制以及该机制是否涉及逆转多药耐药（MDR）的问题尚不完全清楚。本文描述了对 APS 对阿霉素耐药的 H22 肝癌细胞系（H22/ADM）中 P-糖蛋白功能和表达的影响的研究。
方法
用不同浓度的 APS 和/或最常用的化疗药物如环磷酰胺、阿霉素、5-氟尿嘧啶、顺铂、依托泊苷和长春新碱处理 H22/ADM 细胞系。用 MTT 法、流式细胞术、Western blot 和实时定量 RT-PCR 检测化疗药物敏感性、P-糖蛋白功能和表达以及 MDR1mRNA 表达。
结果
APS 单独使用时对体外 H22/ADM 细胞无抗肿瘤活性。然而，它可以增加某些化疗药物如环磷酰胺、阿霉素、5-氟尿嘧啶、顺铂、依托泊苷和长春新碱在 H22/ADM 细胞中的细胞毒性。它呈剂量依赖性。与空白对照组相比，APS 以剂量依赖性方式增加细胞内罗丹明-123 保留并降低 P-糖蛋白外排功能。这些因素在给药后 24 小时、48 小时和 72 小时进行评估。APS 以浓度依赖性方式在 0.8-500mg/L 的最终范围内并在 24-72 小时的时间依赖性方式下调 P-糖蛋白和 MDR1mRNA 表达。
结论
APS 可以增强 H22/ADM 细胞的化疗敏感性。这可能涉及下调 MDR1mRNA 表达、抑制 P-GP 外排泵功能或两者兼而有之，从而降低 MDR1 蛋白的表达。
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