熱治療經由粒線體介導的細胞凋亡機制促進阿黴素的抗癌作用
黃一勝1,2,3,4、張安辰2、陳栢均2、劉秀雯2、蔡德甫1、林宜佳1,4、仇光宇1、陳宏恩1、何肇晏1
1新光醫院 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
Anti-cancer action of hyperthermia in combination with doxorubicin on activating mitochondrial apoptosis
Thomas I-Sheng Hwang1,2,3,4, An-Chen Chang2, Po-Chun Chen2, Hsiu-Wen Liu2, Te-Fu Tsai1, Yi-Chia Lin1,4, Kuang-Yu Chou1, Hung-En Chen1 and Chao-Yen Ho1
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:
Bladder cancer (BC) is the second most common urological tumor in male. Currently, the treatment regimens for bladder cancer are dissatisfied, and the development of alternative therapeutic strategies is urgent. In the previous study, hyperthermia improved the therapeutic efficacy of doxorubicin in cancer cells was examined. However, the mechanism of action mediated by hyperthermia in BC is still unclear. This study aims to analyze whether hyperthermia increases doxorubicin-based chemotherapy in BC cells via the mitochondrial apoptosis pathway.
Materials and methods:
The bladder cancer cells (T24 & 5637) and normal bladder epithelial cell (SV-HUC1) were incubated with doxorubicin/hyperthermia (43℃) for 24 h, 48 h or 7 days. The cell viability and proliferation were assessed by AlamarBlue assay and colony formation assay, respectively. The bladder cancer cells treatment under the conditions described above were subjected to DAPI staining, caspase-3 activity assay, TUNEL assay, and Western blot to confirm the cell apoptosis after combinational treatment of doxorubicin/ hyperthermia. Finally, the molecular mechanism in response to hyperthermia will be measured by Western blot.
Results:
The results showed that hyperthermia enhanced bladder cancer cells' sensitivity to doxorubicin treatment, with a dramatic decrease of IC50 in BC cells. Meanwhile, the colony formation results showed obvious suppression of cell growth in response to the combination treatment of doxorubicin/hyperthermia compared with doxorubicin alone. The cell apoptosis was also dramatically increased after co-treatment of doxorubicin/ hyperthermia, confirming by DAPI staining, caspase-3 activity assay, TUNEL assay, and Western blot. Finally, we evaluated the apoptotic pathway and the data indicated the effects of hyperthermia were exerted by mitochondrial destruction.
Conclusion:
Hyperthermia may serve as a potential adjunctive treatment for bladder cancer. The present study will provide novel insight into discovering a new therapeutic strategy that is ready to use clinically.