Chloroquine induces lysosomal membrane permeability-mediated cell death in bladder cancer cells
Chloroquine induces lysosomal membrane permeability-mediated cell death in bladder cancer cells
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DOI：
10.4103/fjs.fjs_83_17
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发表时间：
2018-07-01
影响因子：
0.4
通讯作者：
Hwang, I-Sheng Thomas
中科院分区：
文献类型：
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作者：
Chen, Hung-En;Lin, Ji-Fan;Hwang, I-Sheng Thomas
背景：氯喹(CQ)与其他化疗药物联合治疗癌症时，被认为是一种有效的佐剂。然而，CQ单次作用对膀胱癌(BC)细胞的影响尚未见报道。用LysoTracker检测溶酶体形态。采用吖啶橙(AO)易位和组织蛋白酶B、D释放的方法检测溶酶体膜通透性(LMP)的变化。Bid、caspase-3和胞浆细胞色素C(Cyto.C)用Western印迹法检测CQ处理细胞中的表达。用胃抑素A和E64d拮抗CQ诱导的LMP。结果：CQ单次作用可诱导BC细胞死亡，胃抑素A和E64d可抑制其作用。LysoTracker染色显示CQ处理细胞的荧光减弱，表明CQ以溶酶体功能为靶标。这进一步得到了更多的AO转位和CatB和CatD释放到细胞质中的支持。裂解出价和胞质细胞数量的增加。C提示线粒体外膜通透性增强，进而导致Caspase3活性升高，进而诱导细胞凋亡。结论：CQ诱导的LMP可促进细胞凋亡，最终导致BC细胞死亡。研究结果首次证明，单一的CQ通过诱导LMP和随后的线粒体膜通透性触发细胞凋亡来对抗BC细胞，使其成为未来BC治疗的潜在治疗方法。
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 (BC) cells have not been investigated.Methods: The growth and viability of CQ-treated BC cells were examined. The lysosomal morphology was detected using LysoTracker. The induction of lysosomal membrane permeability (LMP) was detected by acridine orange (AO) translocation, and cathepsin B and D release. The expression of the bid, caspase-3, and cytosolic cytochrome C (Cyto. C) in CQ-treated cells was detected by the Western blot. The pepstatin A and E64d were used to attenuate CQ-induced LMP.Results: A single dose of CQ treatment induced BC cell death, and attenuated by pepstatin A and E64d. 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 the cytosol. The increased level of cleavage bid and cytosolic Cyto. C indicated mitochondrial outer membrane permeabilization and subsequently leading to apoptosis induction judged by the increased level of activated caspase 3.Conclusion: CQ-induced LMP that enhances apoptosis and ultimately leading to BC cell death. The study results demonstrated for the first time that single CQ treatment against BC cells by inducing LMP and subsequent mitochondria membrane permeability that trigger apoptosis, making it a potential treatment for BC therapy in the future.