抗真菌藥物咪康唑 (Miconazole) 藉由細胞自噬流通量啟動膀胱癌細胞存活的保護機制
蔡德甫1、張安辰2、陳栢均2、劉秀雯2、陳宏恩1、林宜佳1、仇光宇1、林致凡2、黃一勝1,2,3,4
1新光醫院 外科部 泌尿科、2新光醫院 中央研究室、3台北醫學大學 醫學院、4輔仁大學 醫學院
Miconazole, an anti-fungal agent, induced autophagic flux serve as a protective mechanism for the survival of bladder cancer cells
Te-Fu Tsai 1, An-Chen Chang2, Po-Chun Chen2, Hsiu-Wen Liu 2, Hung-En Chen1, Yi-Chia Lin1, Kuang-Yu Chou1, Ji-Fan Lin2, and Thomas I-Sheng Hwang1,3,4,5
Department of Urology, Shin Kong Wu Ho-Su Memorial Hospital1, Central Laboratory, Shin Kong Wu Ho-Su Memorial Hospital2, Department of Urology, Taipei Medical University3, Division of Urology, School of Medicine, Fu-Jen Catholic University4, Taipei, Taiwan.
Background:
Autophagy plays a dual function in cancer progression; autophagy activation can support cancer cell survival or contribute to cell death. In this regard, the development of novel autophagy regulators participating in different autophagy processes can provide information regarding autophagy in cancer biology and its clinical application. Miconazole (MIC), a Food and Drug Administration-approved antifungal drug, has been implicated in oncology research recently. MIC was found to exert antitumor effects in various tumors, including bladder cancer (BC). However, whether it provokes autophagy has never been discussed.
Materials and Methods:
Human bladder cancer cells (T24, 5637) were used to investigate the mechanism of MIC in this study. The Western blot, qPCR and immunofluorescence staining were used to detect the levels of autophagy-related gene expression including LC3-II and p62. The acridine orange, Lysotracker, and cathepsin D staining were used to confirm lysosome formation. The cell viability assay and Annexin/PI staining were performed to monitor apoptosis after MIC exposure.
Results:
Our findings are the first to show that MIC induces autophagy in BC. Based on our in vitro experiments, we found that 1A/1B-light chain 3 (LC3)-II processing and p62 expression were elevated after MIC stimulation through immunofluorescence staining. Besides, adenosine monophosphate-activated protein kinase (AMPK) phosphorylation, which contributes to the autophagy activation, was increased after MIC treatment. Next, bafilomycin A1 (Baf A1), an inhibitor to block autophagosome-lysosome fusion in the late stage of autophagic flux, was used to confirm the effect of autophagy-promoting action after MIC treatment. The result showed that the combination treatment of MIC and Baf A1 significantly improved LC3-II processing, confirming that MIC promoted autophagic flux. The acridine orange, Lysotracker, and cathepsin D staining results revealed that MIC increased lysosome formation, confirming its autophagy-promoting function. Finally, cotreatment of MIC and autophagy inhibitor 3-methyladenine further reduced the cell viability and induced apoptosis in BC cells, proving that MIC provokes protective autophagy in BC cells.
Conclusions:
Our findings approved MIC as a novel autophagy inducer, which can be used to determine the role of protective autophagy in cancer treatment.