Rotem Ronit, Heyfets Alina, Fingrut Orit, Blickstein Dorit, Shaklai Mati, Flescher Eliezer
Department of Human Microbiology, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel.
Cancer Res. 2005 Mar 1;65(5):1984-93. doi: 10.1158/0008-5472.CAN-04-3091.
We reported previously that jasmonates can kill human cancer cells. Many chemotherapeutic drugs induce mitochondrial membrane permeability transition, membrane depolarization, osmotic swelling, and release of cytochrome c, involving the opening of the permeability transition pore complex (PTPC). Because jasmonates exert their cytotoxic effects independent of transcription, translation, and p53 expression, we hypothesized that these compounds may act directly on mitochondria. Mitochondrial membrane depolarization was determined by flow cytometry, and cytochrome c release by Western blotting. Mitochondria were isolated by mechanical lysis and differential centrifugation. Cytotoxicity was measured by a tetrazolium-based assay, and mitochondrial swelling by spectrophotometry. Jasmonates induced membrane depolarization and cytochrome c release in intact human cancer cell lines. Jasmonates induced swelling in mitochondria isolated from Hep 3B hepatoma cells, but not in mitochondria isolated from 3T3 nontransformed cells or from normal lymphocytes, in a PTPC-mediated manner. Methyl jasmonate induced the release of cytochrome c from mitochondria isolated from cancer cell lines in a PTPC-mediated manner, but not from mitochondria isolated from normal lymphocytes. A correlation was found between cytotoxicity of methyl jasmonate and the percentage of leukemic cells in the blood of patients with chronic lymphocytic leukemia (CLL). Jasmonates induced membrane depolarization in CLL cells, and swelling and release of cytochrome c in mitochondria isolated from these cells. In conclusion, jasmonates act directly on mitochondria derived from cancer cells in a PTPC-mediated manner, and could therefore bypass premitochondrial apoptotic blocks. Jasmonates are promising candidates for the treatment of CLL and other types of cancer.
我们之前报道过茉莉酸酯能杀死人类癌细胞。许多化疗药物会诱导线粒体膜通透性转换、膜去极化、渗透性肿胀以及细胞色素c的释放,这涉及通透性转换孔复合物(PTPC)的开放。由于茉莉酸酯发挥其细胞毒性作用独立于转录、翻译和p53表达,我们推测这些化合物可能直接作用于线粒体。通过流式细胞术测定线粒体膜去极化,通过蛋白质免疫印迹法测定细胞色素c的释放。通过机械裂解和差速离心分离线粒体。通过基于四氮唑的测定法测量细胞毒性,通过分光光度法测量线粒体肿胀。茉莉酸酯在完整的人类癌细胞系中诱导膜去极化和细胞色素c释放。茉莉酸酯以PTPC介导的方式诱导从Hep 3B肝癌细胞分离的线粒体肿胀,但不诱导从3T3未转化细胞或正常淋巴细胞分离的线粒体肿胀。茉莉酸甲酯以PTPC介导的方式诱导从癌细胞系分离的线粒体释放细胞色素c,但不诱导从正常淋巴细胞分离的线粒体释放。发现茉莉酸甲酯的细胞毒性与慢性淋巴细胞白血病(CLL)患者血液中白血病细胞的百分比之间存在相关性。茉莉酸酯在CLL细胞中诱导膜去极化,并在从这些细胞分离的线粒体中诱导肿胀和细胞色素c释放。总之,茉莉酸酯以PTPC介导的方式直接作用于癌细胞来源的线粒体,因此可以绕过线粒体前凋亡阻滞。茉莉酸酯是治疗CLL和其他类型癌症的有前景的候选药物。