激活磷酸腺苷活化蛋白激酶（AMPK）信号通路在伊曲康唑诱导的抗食管癌活性中的作用和机制研究

食管癌是常见恶性肿瘤之一，预实验结果显示抗真菌药物伊曲康唑（Itra）能显著抑制食管癌细胞存活、增殖，并诱导癌细胞自噬。机制方面Itra诱导神经酰胺合成而激活酸腺苷活化蛋白激酶（AMPK），并诱导生长因子受体EGFR/PDGFR降解，同时阻断mTOR和Hh-Gli1信号活化；而AMPK抑制剂及AMPK-shRNA逆转上述作用。我们提出神经酰胺介导的AMPK活化是Itra抗食管癌作用的核心机制。本课题中，申请者将利用分子、细胞生物学及裸鼠荷瘤等方法，体内外观察Itra的抗食管癌作用。重点研究Itra抗癌的分子机制，阐明AMPK信号通路在其中的作用。并调节癌细胞内神经酰胺含量和AMPK活化水平，达到增敏Itra抗癌的目的。最后检测AMPKα1在食管癌组织中的表达及活化水平，并分析其与肿瘤各项临床指标的相关性。旨在明确Itra抗食管癌作用和分子机制，为开发Itra治疗食管癌提供理论和实验依据。

The esophageal carcinoma is one of the most malignant human tumors with extremely poor prognosis in China and around the world. Our preliminary results found that low concentrations of itraconazole (Itra, at μg/ml), the anti-fungi drug, induced significant growth inhibition and autophagic cell death in cultured esophageal carcinoma cells, which was associated with AMPK signaling activation, ceramide production, mTOR complex 1 (mTORC1) inhibition as well as EGFR/PDGFR degradation and Hedgehog(Hh)-Gli1 signaling in-activation. We propose that Itra-induced anti-esophageal carcinoma efficiency requires ceramide-dependent AMPK signaling activation. The aim of this proposal is to investigate the anti- esophageal carcinoma effect of Itra both in vivo and in vitro, and to study the underlying mechanisms by focusing on the AMP-activated protein kinase (AMPK) signaling. By using multiple methods, we are set to manipulate ceramide-AMPK signaling to increase Itra's efficiency. Finally, we will test the expression and activation of AMPKα1 in human esophageal carcinoma tissues, and analyze its relationship with tumor staging, patients' prognosis and other clinical outcomes. This study suggests that AMPK signaling might be the key determinant of Itra's response in esophageal carcinoma and probably other cancer cells. The results of this study will provide theoretical and experimental evidence for using Itra as a novel anti-esophageal carcinoma agent.