Epithelial to mesenchymal transition contributes to drug resistance in pancreatic cancer.
Epithelial to mesenchymal transition contributes to drug resistance in pancreatic cancer.
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DOI：
10.1158/0008-5472.can-08-2819
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发表时间：
2009-07-15
期刊：
影响因子：
11.2
通讯作者：
Choi W
中科院分区：
文献类型：
--
作者：
Arumugam T;Ramachandran V;Fournier KF;Wang H;Marquis L;Abbruzzese JL;Gallick GE;Logsdon CD;McConkey DJ;Choi W
为了改善胰腺癌患者的预后，需要更好地了解耐药机制。在这里，我们描述了胰腺癌细胞对三种具有不同作用机制的常规化疗药物（吉西他滨、5-氟尿嘧啶和顺铂）的敏感性和耐药模式。4例（L3.6pl、BxPC-3、CFPAC-1、SU86.86）对3种药物均敏感，5例（PANC-1、Hs766T、AsPC-1、MIAPaCa-2、Mpanc96）对3种药物均耐药（GI50（50%生长抑制））。基因表达谱和无监督分层聚类分析显示，敏感细胞和耐药细胞形成了两个不同的组，在特定基因的表达上存在差异，包括“上皮-间质转化”（EMT）的几个特征。有趣的是，在基因表达数据中观察到E-cadherin与其转录抑制因子Zeb-1呈负相关，并通过实时PCR证实。5个新的胰腺癌细胞系的独立验证实验证实，E-cadherin和Zeb-1的负相关与对吉西他滨、5-氟尿嘧啶和顺铂的耐药密切相关。在间充质细胞系中沉默Zeb-1不仅增加了E-cadherin的表达，还增加了EVA1和MAL2等其他上皮标志物的表达，并恢复了药物敏感性。重要的是，E-cadherin和Zeb-1在原发肿瘤中的免疫组化分析证实了这两种蛋白的表达是相互排斥的（p=0.012）。因此，我们的研究结果表明，Zeb-1和其他EMT调节因子可能在人胰腺癌细胞中维持耐药，抑制Zeb-1和逆转EMT的治疗策略应该得到评估。
A better understanding of drug resistance mechanisms is required to improve outcomes in patients with pancreatic cancer. Here we characterized patterns of sensitivity and resistance to three conventional chemotherapeutic agents with divergent mechanisms of action (gemcitabine, 5-fluorouracil, and cisplatin) in pancreatic cancer cells. Four (L3.6pl, BxPC-3, CFPAC-1, SU86.86) were sensitive and five (PANC-1, Hs766T, AsPC-1, MIAPaCa-2, Mpanc96) were resistant to all 3 agents based on GI50 (50% growth inhibition). Gene expression profiling and unsupervised hierarchical clustering revealed that the sensitive and resistant cells formed two distinct groups and differed in expression of specific genes including several features of “epithelial-mesenchymal transition” (EMT). Interestingly, an inverse correlation between E-cadherin and its transcriptional suppressor, Zeb-1 was observed in the gene expression data and was confirmed by real time PCR. Independent validation experiment using 5 new pancreatic cancer cell lines confirmed that an inverse correlation between E-cadherin and Zeb-1 correlated closely with resistance to gemcitabine, 5-fluorouracil, and cisplatin. Silencing Zeb-1 in the mesenchymal lines not only increased the expression of E-cadherin but also other epithelial markers such as EVA1 and MAL2 and restored drug sensitivity. Importantly, immunohistochemical analysis of E-cadherin and Zeb-1 in primary tumors confirmed that expression of the two proteins was mutually exclusive (p=0.012). Therefore, our results suggest that Zeb-1 and other regulators of EMT may maintain drug resistance in human pancreatic cancer cells, and therapeutic strategies to inhibit Zeb-1 and reverse EMT should be evaluated.