Chloroquine 引發膀胱癌細胞之溶酶體破壞並導致細胞凋亡
黃一勝1,2,3、林致凡4,、林宜佳1,3,、蔡德甫1,3,、陳宏恩1、仇光宇1,3
1新光醫院 外科部 泌尿科; 2台北醫學大學醫學系 泌尿學科; 3輔仁大學醫學系 泌尿學科; 4新光醫院中央實驗室
Chloroquine induces lysosomal disruptions and apoptotic cell death in bladder cancer cells
Thomas I-Sheng Hwang1,2,3, Ji-Fan Lin4, Yi-Chia Lin1,3, Te-Fu Tsai 1,3,Hung-En Chen1, Kuang-Yu Chou1,3
1Department of Urology, Shin Kong Wu Ho-Su Memorial Hospital, Department of Urology,
2Taipei Medical University, Division of Urology, 3School of Medicine, Fu-Jen Catholic University, 4Central Laboratory, Shin Kong Wu Ho-Su Memorial Hospital, Taipei, Taiwan.
Background: Chloroquine (CQ) is recognized as a potent adjuvant when combined with other chemotherapies to treat cancers. However, the effects of a single treatment of CQ on bladder cancer have not been investigated.
Methods: The growth and viability of CQ-treated cells were examined by Cytosmart system and WST-1. The lysosomal morphology was detected by LysoTracker. The induction of lysosomal membrane permeability (LMP) was detected by acridine orange translocation,and cathepsin B (CatB) and D(CatD) release. The expression of bid, caspase-3, and cytosolic cytochrome C in CQ-treated cells was detected by Western blot. The pepstatinA and E64d were used to attenuate CQ-induced LMP.
Results: A single dose of CQ treatment induced bladder cancer cell death, and attenuated by pepstatin A and E64d, protease inhibitors. The diminishing of fluorescent in CQ-treated cells stained with LysoTracker, suggesting that CQ targets lysosomal functions. This was further supported by increased AO translocation, and the releasing of CatB and CatD into cytosol. The increased level of cleavage bid and cytosolic cytochrome c indicated mitochondrial outer membrane permeabilization (MOMP), and subsequently leading to apoptosis induction judged by the increased level of caspase 3.
Conclusions: CQ induced LMP that amplifies apoptosis and ultimately leading to bladder cancer cell death.