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下调 miR-214 通过靶向 Twist 促进肝内胆管癌转移。

Down-regulation of miR-214 contributes to intrahepatic cholangiocarcinoma metastasis by targeting Twist.

机构信息

Department of Biliary Truct Surgery I, Eastern Hepatobiliary Hospital, Secondary Military Medicine University, Shanghai, China.

出版信息

FEBS J. 2012 Jul;279(13):2393-8. doi: 10.1111/j.1742-4658.2012.08618.x.

Abstract

miRNAs play an important role in many human diseases, including cancer metastasis. However, the mechanisms by which miRNAs regulate intrahepatic cholangiocarcinoma metastasis remain poorly understood. In the present study, we assayed the expression level of miR-214 in intrahepatic cholangiocarcinoma tissues by real-time PCR, and defined the target gene and biological function by luciferase reporter assay and Western blot analysis. We found that the miR-214 levels were remarkably decreased in metastatic intrahepatic cholangiocarcinoma tissues compared to non-metastatic tissues. Inhibition of miR-214 levels by its inhibitor promoted metastasis of human intrahepatic cholangiocarcinoma cell. We further demonstrated that down-regulation of miR-214 increased the transcript levels of the epithelial-mesenchymal transition-associated gene Twist, and then decreased E-cadherin levels. We confirmed that down-regulation of miR-214 promoted the epithelial-mesenchymal transition by directly targeting the Twist gene. These results suggest an important role for miR-214 in regulating metastasis of intrahepatic cholangiocarcinoma, and potential application of miR-214 in intrahepatic cholangiocarcinoma therapy.

摘要

miRNAs 在许多人类疾病中发挥着重要作用,包括癌症转移。然而,miRNAs 调节肝内胆管癌转移的机制仍知之甚少。在本研究中,我们通过实时 PCR 检测了 miR-214 在肝内胆管癌组织中的表达水平,并通过荧光素酶报告基因检测和 Western blot 分析确定了靶基因和生物学功能。我们发现,转移性肝内胆管癌组织中的 miR-214 水平明显低于非转移性组织。通过其抑制剂抑制 miR-214 水平可促进人肝内胆管癌细胞的转移。我们进一步证明,下调 miR-214 可增加上皮-间充质转化相关基因 Twist 的转录水平,从而降低 E-钙黏蛋白水平。我们证实下调 miR-214 通过直接靶向 Twist 基因促进上皮-间充质转化。这些结果表明 miR-214 在调节肝内胆管癌转移中具有重要作用,并为 miR-214 在肝内胆管癌治疗中的潜在应用提供了依据。

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