Miconazole在膀胱癌抑癌效果及凋亡機制

膀胱癌為一種常見且極具侵略性的惡性腫瘤，早期症狀不易察覺，所以發現時都已發生轉移。在台灣，根據行政院衛生署2011年癌症登記年報統計，膀胱癌占所有癌症發生率排名於男性為第九、女性為第十六位。抗黴菌藥物Miconazole在之前的研究中已經證實具抑制乳腺癌、大腸癌和人類骨肉瘤等癌細胞，而膀胱癌尚未有研究報導。因此，此篇研究目的為探討膀胱癌與Miconazole間的作用機制。實驗結果顯示，利用MTT assay確認Miconazole可抑制膀胱癌細胞TSGH8301的生長，藉由流式細胞儀證實藥物對細胞週期產生影響，也藉由DNA fragmentation、Annexin-FITC / PI 染色等實驗證實癌細胞走向凋亡，並由JC-1染色發現與粒線體路徑有關；更進一步以西方墨點法確認，發現p53、p21和p27蛋白量增加，伴隨著Cyclin D1、Cyclin E1、CDK2和CDK4蛋白量減少，Caspase-3、Caspase-9和cleaved PRAR蛋白增加，以及細胞質內Cytochrome c和SMAC蛋白的增加。這些結果顯示了Miconazole可以抑制膀胱癌細胞TSGH8301的生長並誘導細胞走向粒線體凋亡路徑，這意味著Miconazole具有潛力作為治療人類膀胱癌的藥物。

Bladder cancer is a very frequent and aggressive malignant tumor. Bladder cancer is difficult to detect at early stages, and is usually at the late stages when diagnosed. In Taiwan, bladder cancer is the ninth most frequent malignancy diagnosed in men and the 16th in women based on a 2011 report from the Department of Health, Republic of China. Miconazole (MIC) is an imidazole antifungal agent, in recently reports, MIC have been shown to inhibit a variety of cancers, for example, breast cancer, mammary cancer and osteosarcoma. But there are not any research about bladder cancer. The aim of this study was to determine the effects of MIC on the growth inhibition and apoptosis of human bladder cancer cell. In the study showed that MIC elicited cytotoxic effects on human TSGH 8301 bladder as determined by MTT assay. The drug also impact on cell cycle confirmed by flow cytometry. MIC could induced DNA fragmentation formation and Annexin V-FITC / PI stained, toward to mitochondria-mediated cell apoptosis pathway as determined by JC-1 stain. Western blot analysis revealed that the increases of p21 and p27 protein levels, alone with the decrease of Cyclin E1, CDK2 and CDK4 expressed in MIC-treated TSGH-8301 cells, activation of caspase-3/-9, and cleavage PARP, and cytochrome c and SMAC protein increased from cytoplasm. These results indicate that the mitochondrial pathway is involved in MIC-induced growth inhibition and apoptosis of human bladder cancer, suggesting that MIC may be a potential anti-bladder cancer agent in human.