Targeted delivery of doxorubicin through conjugation with EGF receptor-binding peptide overcomes drug resistance in human colon cancer cells
Targeted delivery of doxorubicin through conjugation with EGF receptor-binding peptide overcomes drug resistance in human colon cancer cells
复制标题
通过与 EGF 受体结合肽缀合靶向递送阿霉素克服人结肠癌细胞的耐药性
DOI：
10.1111/bph.12055
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发表时间：
2013-04-01
影响因子：
7.3
通讯作者：
Huang, Zebo
中科院分区：
文献类型：
--
作者：
Ai, Shibin;Jia, Tao;Huang, Zebo
背景与目的多柔比星（DOX）诱导的多药耐药及非特异性毒性反应限制了以DOX为基础的化疗。最近，我们证明了DOX与EGF受体结合肽（DOX-EBP）结合，在靶向EGF受体过表达肿瘤时，具有增强的抗癌功效和降低的全身毒性。在此，我们研究了DOX-EBP是否能够克服耐药性和潜在的分子机制。实验方法通过逐步增加DOX浓度，从非抗性SW 480细胞衍生DOX抗性SW 480/DOX细胞，并且通过Western印迹证实DOX诱导的P-糖蛋白过表达。分别采用MTT法和荧光显微镜评价药物的细胞毒性和细胞内分布。EGF受体介导的内吞作用在EGF受体和内吞抑制试验中测定。通过HPLC测定肿瘤细胞和鼠异种移植物中的药物蓄积。关键结果DOX-EBP在SW 480/DOX细胞中的细胞毒性和蓄积几乎与SW 480细胞相同，但游离DOX的细胞毒性和蓄积降低。DOX-EBP积累被EGF受体和内吞作用的抑制剂阻止，表明EGF受体介导内吞摄取。DOX-EBP在小鼠体内的肿瘤蓄积显著高于游离DOX，并且DOX-EBP的水平在DOX耐药和非耐药肿瘤组织中相似。重要的是，DOX-EBP，而不是游离DOX，在敏感和耐药模型中有效抑制实体瘤生长并提高存活率。结论：DOX-EBP能克服肿瘤细胞对DOX的耐药性，提高体内抗肿瘤疗效。因此，它有可能成为治疗耐药性癌症的有效治疗剂。
Background and Purpose Induction of multidrug resistance by doxorubicin (DOX), together with non-specific toxicities, has restricted DOX-based chemotherapy. Recently, we demonstrated that DOX conjugated with an EGF receptor-binding peptide (DOX-EBP) had enhanced anticancer efficacy and reduced systemic toxicity when targeting EGF receptor-overexpressing tumours. Here we investigated whether DOX-EBP is able to overcome drug resistance and the underlying molecular mechanisms. Experimental Approach DOX-resistant SW480/DOX cells were derived from non-resistant SW480 cells by stepwise exposure to increasing concentrations of DOX, and P-glycoprotein overexpression induced by DOX was confirmed by Western blotting. Cytotoxicity and intracellular distribution of drugs were evaluated by MTT assay and fluorescence microscopy respectively. EGF receptor-mediated endocytosis was determined in EGF receptor and endocytosis inhibition assays. Drug accumulation in tumour cells and murine xenografts was determined by HPLC. Key Results The cytotoxicity and accumulation of DOX-EBP in SW480/DOX cells were almost the same as in SW480 cells, but those of free DOX were reduced. DOX-EBP accumulation was prevented by inhibitors of both EGF receptors and endocytosis, suggesting EGF receptors mediate endocytotic uptake. Tumour accumulation of DOX-EBP was significantly higher than free DOX in mice, and the levels of DOX-EBP were similar in DOX-resistant and non-resistant tumour tissues. Importantly, DOX-EBP, but not free DOX, was effective at inhibiting solid tumour growth and increased survival rate in both sensitive and resistant models. Conclusion and Implications DOX-EBP can overcome DOX resistance of tumour cells and increase in vivo antitumour efficacy. Therefore, it has the potential to be a potent therapeutic agent for treating drug-resistant cancers.