P-糖蛋白抗体功能化碳纳米管克服人白血病细胞的多药耐药性。

P-glycoprotein antibody functionalized carbon nanotube overcomes the multidrug resistance of human leukemia cells.

机构信息

National Chromatographic R&A Center, CAS Key Laboratory of Separation Sciences for Analytical Chemistry,Dalian Institute of Chemical Physics, Chinese Academy of Sciences (CAS), Dalian 116023, China.

出版信息

ACS Nano. 2010 Mar 23;4(3):1399-408. doi: 10.1021/nn9011225.

DOI:10.1021/nn9011225

PMID:20148593

Abstract

Multidrug resistance (MDR), which is related to cancer chemotherapy, tumor stem cells, and tumor metastasis, is a huge obstacle for the effective cancer therapy. One of the underlying mechanisms of MDR is the increased efflux of anticancer drugs by overexpressed P-glycoprotein (P-gp) of multidrug resistant cells. In this work, the antibody of P-gp (anti-P-gp) functionalized water-soluble single-walled carbon nanotubes (Ap-SWNTs) loaded with doxorubicin (Dox), Dox/Ap-SWNTs, were synthesized for challenging the MDR of K562 human leukemia cells. The resulting Ap-SWNTs could not only specifically recognize the multidrug resistant human leukemia cells (K562R), but also demonstrate the effective loading and controllable release performance for Dox toward the target K562R cells by exposing to near-infrared radiation (NIR). The recognition capability of Ap-SWNTs toward the K562R cells was confirmed by flow cytometry (FCM) and confocal laser scanning microscopy (CLSM). The binding affinity of Ap-SWNTs toward drug-resistant K562R cells was ca. 23-fold higher than that toward drug-sensitive K562S cells. Additionally, CLSM indicated that Ap-SWNTs could specifically localize on the cell membrane of K562R cells and the fluorescence of Dox in K562R cells could be significantly enhanced after the employment of Ap-SWNTs as carrier. Moreover, the composite of Dox and Ap-SWNTs (Dox/Ap-SWNTs) expressed 2.4-fold higher cytotoxicity and showed the significant cell proliferation suppression toward K562R leukemia cells (p < 0.05) as compared with free Dox which is popularly employed in clinic trials. These results suggest that the Ap-SWNTs are the promising drug delivery vehicle for overcoming the MDR induced by the overexpression of P-gp on cell membrane. Ap-SWNTs loaded with drug molecules could be used to suppress the proliferation of multidrug resistant cells, destroy the tumor stem cells, and inhibit the metastasis of tumor.

摘要

多药耐药性（MDR）与癌症化疗、肿瘤干细胞和肿瘤转移有关，是癌症有效治疗的巨大障碍。MDR 的一个潜在机制是多药耐药细胞中过表达的 P 糖蛋白（P-gp）增加了抗癌药物的外排。在这项工作中，合成了载有阿霉素（Dox）的 P-gp（抗-P-gp）功能化的水溶性单壁碳纳米管（Ap-SWNTs）负载药物（Dox/Ap-SWNTs），用于挑战人白血病 K562 细胞的多药耐药性。结果表明，Ap-SWNTs 不仅可以特异性识别多药耐药的人白血病细胞（K562R），而且通过暴露于近红外辐射（NIR），对靶标 K562R 细胞表现出有效的载药和可控释放性能。通过流式细胞术（FCM）和共聚焦激光扫描显微镜（CLSM）证实了 Ap-SWNTs 对 K562R 细胞的识别能力。Ap-SWNTs 对耐药 K562R 细胞的结合亲和力约为对敏感 K562S 细胞的 23 倍。此外，CLSM 表明 Ap-SWNTs 可以特异性定位于 K562R 细胞的细胞膜上，并且在将 Ap-SWNTs 用作载体后，K562R 细胞中的 Dox 荧光可以显著增强。此外，与广泛用于临床试验的游离 Dox 相比，Dox/Ap-SWNTs （Dox/Ap-SWNTs）的组合表现出 2.4 倍更高的细胞毒性，并对 K562R 白血病细胞表现出显著的细胞增殖抑制作用（p <0.05）。这些结果表明，Ap-SWNTs 是克服细胞膜上过表达 P-gp 引起的多药耐药性的有前途的药物传递载体。负载药物分子的 Ap-SWNTs 可用于抑制多药耐药细胞的增殖，破坏肿瘤干细胞并抑制肿瘤转移。