Curcumin mediated down-regulation of αVβ3 integrin and up-regulation of pyruvate dehydrogenase kinase 4 (PDK4) in Erlotinib resistant SW480 colon cancer cells
Curcumin mediated down-regulation of αVβ3 integrin and up-regulation of pyruvate dehydrogenase kinase 4 (PDK4) in Erlotinib resistant SW480 colon cancer cells
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DOI：
10.1002/ptr.5984
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发表时间：
2018-02-01
影响因子：
7.2
通讯作者：
Fathi, Mojtaba
中科院分区：
文献类型：
--
作者：
Javadi, Samira;Rostamizadeh, Kobra;Fathi, Mojtaba
厄洛替尼是一种有效的，选择性的，口服活性的表皮生长因子受体抑制剂，但化疗期间厄洛替尼耐药性的发展可能导致治疗失败。为了阐明厄洛替尼耐药途径，本研究调查了使用姜黄素和厄洛替尼负载纳米颗粒的联合治疗对厄洛替尼耐药SW 480结肠癌细胞系中α（V）β（3）整联蛋白和丙酮酸脱氢酶激酶4（PDK 4）表达的影响。通过长期暴露于厄洛替尼产生了厄洛替尼抗性SW 480结肠癌细胞系。使用单步纳米沉淀法提供姜黄素负载的甲氧基聚乙二醇聚己内酯（cur/mPEG-PCL）和厄洛替尼负载的mPEG-PCL（erl/mPEG-PCL）胶束，并用作抗性SW 480癌细胞的联合治疗。之后，通过半定量逆转录-聚合酶链反应测定PDK 4、α v和β 3 mRNA的基因表达水平。使用酶联免疫吸附测定法评价整个α（V）β（3）整联蛋白的蛋白水平。在SW 480细胞系中，厄洛替尼对非耐药细胞和耐药细胞的IC 50分别为87.6 +/- 1.2 nM和19.1 +/- 0.14 μ M，姜黄素对非耐药细胞和耐药细胞的IC 50分别为21.8和30 μ M。尽管PDK 4表达在耐药和非耐药细胞中没有显著差异，其表达上调通过剂量为3 μ M的cur/mPEG-PCL和剂量为5 μ M的erl/mPEG-PCL的联合治疗，在抗性细胞中产生的α（V）β（3）mRNA和整个α（V）β（3）的蛋白质水平增加了1.4倍SW 480耐药细胞中整合素水平显著高于非耐药细胞。在治疗方面，6 μ M cur/mPEG-PCL和5 μ M erl/mPEG-PCL的组合与非抗性细胞相比，在抗性细胞中下调β（3）基因表达6.6倍。在蛋白质水平，3 μ M-cur/mPEG-PCL和10 μ M erl/mPEG-PCL的组合减少了抗性细胞中的α（V）β（3）蛋白。结果表明cur/mPEG-PCL和erl/mPEG-PCL联合治疗可降低耐药结肠癌细胞中α（V）β（3）整合素的表达，并增加PDK 4基因的表达，这可能影响耐药信号通路。
Erlotinib is a potent, selective, and orally active inhibitor of the epidermal growth factor receptor, but the development of erlotinib resistance during chemotherapy can lead to treatment failure. To shed light on the erlotinib-resistant pathway, this study investigated the effect of combination therapy using curcumin- and erlotinib-loaded nanoparticles on the expression of alpha(V)beta(3) integrin and pyruvate dehydrogenase kinase 4 (PDK4) in an erlotinib-resistant SW480 colon cancer cell line. An erlotinib-resistant SW480 colon cancer cell line was produced by long-term exposure to erlotinib. Curcumin-loaded Methoxy poly ethylene glycol Poly caprolactone (cur/mPEG-PCL) and erlotinib-loaded mPEG-PCL (erl/mPEG-PCL) micelles were provided using a single step nanoprecipitation method and used as combination therapy of resistant SW480 cancer cells. After that, gene expression levels of PDK4, alpha v, and beta 3 mRNA were determined by the semiquantitative reverse transcription-polymerase chain reaction. Protein levels of whole alpha(V)beta(3) integrin were evaluated using the enzyme-linked immunosorbent assay method. In SW480 cell line, the IC50 of nonresistant and resistant cells was 87.6 +/- 1.2 nM and 19.1 +/- 0.14 mu M, for erlotinib and it was about 21.8 and 30 mu M for curcumin, respectively. Although PDK4 expression was not significantly different in resistant and nonresistant cells, its expression was up regulated (1.4 fold) in resistant cells by a combination therapy of cur/mPEG-PCL at a dose of 3 mu M and erl/mPEG-PCL at a dose of 5 mu M.beta(3) mRNA and the protein level of whole alpha(V)beta(3) integrin was significantly higher in resistant SW480 cells as compared with those in nonresistant cells. In terms of treatment, a combination of 6-mu M cur/mPEG-PCL and 5-mu M erl/mPEG-PCL down regulated beta(3) gene expression 6.6-fold in resistant cells as compared with nonresistant cells. At the protein level, a combination of 3-mu M-cur/mPEG-PCL and 10-mu M erl/mPEG-PCL reduced alpha(V)beta(3) protein in resistant cells. The results indicated that combination therapy using cur/mPEG-PCL and erl/mPEG-PCL could decrease alpha(V)beta(3) integrin expression and increase PDK4 gene expression in resistant colon cancer cells, which may have effects on drug resistance signaling pathways.