姜黄素通过调节 P-糖蛋白对 MNNG/HOS 人骨肉瘤细胞在体、内外多药耐药的逆转作用。

Reversion effects of curcumin on multidrug resistance of MNNG/HOS human osteosarcoma cells in vitro and in vivo through regulation of P-glycoprotein.

机构信息

Department of Orthopedics, Qilu Hospital of Shandong University, Jinan, Shandong 250012, China.

出版信息

Chin Med J (Engl). 2013 Nov;126(21):4116-23.

Abstract

BACKGROUND

P-glycoprotein (P-gp) encoded by ATP-binding cassette sub-family B member 1 (ABCB1) gene is a kind of ATP-dependent drug transporter, which plays important roles in multidrug resistance (MDR) of human cancers, such as osteosarcoma. Curcumin is a natural phenolic coloring compound originating from the rhizomes of Curcuma longa, which is proved to possess antitumor biological activities including reversion of MDR. However, the effect and molecular mechanisms of curcumin to osteosarcoma MDR remain unclear.

METHODS

We established a human osteosarcoma drug-resistant cell line MNNG/HOS/MTX by pulse exposure to methotrexate (MTX) and verified that the new cell lines were cross-resistant to other anticancer agents. Then, according to the cytotoxicity assay, we reversed MDR of MNNG/HOS/MTX by 30 µmol/L curcumin, and detected the mechanisms of curcumin reversing MDR through Real-time PCR, Western blotting assay, and Rhodamine123 (Rh123) transport test. Finally, we evaluated the effect of curcumin reversing MDR in vivo by MNNG/HOS/MTX cells xenograft-nude mice model.

RESULTS

MNNG/HOS/MTX was proved to be a human osteosarcoma MDR cell line. MTT tumor chemosensitivity test indicates that 30 µmol/L curcumin attenuates the half maximal inhibitory concentration (IC50) and resistance index (RI) to MTX, diamminedichloroplatinum (DDP), adriamycin (ADM), ifosfamide (IFO), and epirubicin (EPI) in MNNG/HOS/MTX cells (P < 0.05). Real-time PCR and Western blotting assays demonstrated that curcumin down-regulated P-gp expression of MNNG/HOS/MTX cells. Rh123 transport test showed that curcumin inhibited the transport function of P-gp in vitro. In vivo studies showed that curcumin displayed the features of sensitizing antitumor drugs and inhibiting the proliferation, invasion, and metastasis of osteosarcoma MDR cells.

CONCLUSION

Down-regulation of P-gp and inhibition of the function of P-gp efflux pump may contribute to MDR reversion induced by curcumin in vitro and in vivo.

摘要

背景

P-糖蛋白（P-gp）由 ATP 结合盒亚家族 B 成员 1（ABCB1）基因编码，是一种 ATP 依赖性药物转运体，在人类癌症的多药耐药（MDR）中发挥重要作用，如骨肉瘤。姜黄素是一种天然酚类着色化合物，来源于姜黄的根茎，已被证明具有抗肿瘤的生物活性，包括逆转 MDR。然而，姜黄素对骨肉瘤 MDR 的作用和分子机制尚不清楚。

方法

我们通过脉冲暴露于甲氨蝶呤（MTX）建立了人骨肉瘤耐药细胞系 MNNG/HOS/MTX，并验证了新细胞系对其他抗癌药物具有交叉耐药性。然后，根据细胞毒性测定，我们用 30µmol/L 姜黄素逆转 MNNG/HOS/MTX 的 MDR，并通过实时 PCR、Western blot 检测和罗丹明 123（Rh123）转运试验检测姜黄素逆转 MDR 的机制。最后，我们通过 MNNG/HOS/MTX 细胞异种移植裸鼠模型评价姜黄素逆转 MDR 的体内效果。

结果

MNNG/HOS/MTX 被证明是人骨肉瘤 MDR 细胞系。MTT 肿瘤药敏试验表明，30µmol/L 姜黄素降低了 MNNG/HOS/MTX 细胞对 MTX、顺铂（DDP）、阿霉素（ADM）、异环磷酰胺（IFO）和表柔比星（EPI）的半最大抑制浓度（IC50）和耐药指数（RI）（P<0.05）。实时 PCR 和 Western blot 检测表明，姜黄素下调了 MNNG/HOS/MTX 细胞的 P-gp 表达。Rh123 转运试验表明，姜黄素抑制了 P-gp 在体外的转运功能。体内研究表明，姜黄素表现出增敏抗肿瘤药物和抑制骨肉瘤 MDR 细胞增殖、侵袭和转移的特征。