Special Class of Healthcare, Industry Management, Central Taiwan University of Science and Technology, Taichung, Taiwan, ROC.
Int J Oncol. 2010 Nov;37(5):1243-50. doi: 10.3892/ijo_00000775.
Cantharidin has shown potent anticancer activities on many types of human cancer cells. This study was performed to elucidate whether mitochondria and caspases are involved in the modulation of apoptosis and cell cycle arrest by cantharidin in human bladder cancer cells. The effect of cantharidin on cell cycle arrest, apoptosis, caspases, reactive oxygen species (ROS) and mitochondrial membrane potential (ΔΨ(m)) were measured by flow cytometry, and the levels of apoptosis-associated proteins and its regulatory molecules were studied by Western blotting. Cantharidin-induced apoptosis and DNA damage was determined by flow cytometric analysis, DAPI staining and Comet assay. After cantharidin treatment, the active forms of caspase-3, -8 and -9 were promoted. Cantharidin-induced apoptosis was associated with enhanced ROS and Ca(2+) generations, caused DNA damage, decreased the levels of ΔΨ(m) and promoted Endo G and AIF released from mitochondria. Cantharidin-induced G0/G1 arrest was associated with a marked decrease in the protein expressions of cyclin E and Cdc25c but promoted the levels of p21 and p-p53. Cantharidin-induced apoptosis was accompanied with up-regulation of the protein expression of Bax and PARP, but down-regulation of the protein levels of Bcl-2, resulting in dysfunction of mitochondria then led to Endo G and AIF release for causing induction of apoptosis.
斑蝥素在多种人类癌细胞中显示出强大的抗癌活性。本研究旨在阐明线粒体和胱天蛋白酶是否参与斑蝥素对人膀胱癌细胞凋亡和细胞周期阻滞的调节。通过流式细胞术测量斑蝥素对细胞周期阻滞、凋亡、胱天蛋白酶、活性氧（ROS）和线粒体膜电位（ΔΨ（m））的影响，并通过 Western blot 研究凋亡相关蛋白及其调节分子的水平。通过流式细胞术分析、DAPI 染色和彗星试验确定斑蝥素诱导的细胞凋亡和 DNA 损伤。斑蝥素处理后，促进了 caspase-3、-8 和 -9 的活性形式的产生。斑蝥素诱导的细胞凋亡与增强的 ROS 和 Ca（2+）生成、DNA 损伤、ΔΨ（m）水平降低以及促进 Endo G 和 AIF 从线粒体释放有关。斑蝥素诱导的 G0/G1 期阻滞与细胞周期蛋白 E 和 Cdc25c 的蛋白表达明显减少有关，但促进了 p21 和 p-p53 的水平。斑蝥素诱导的细胞凋亡伴随着 Bax 和 PARP 蛋白表达的上调，但 Bcl-2 蛋白水平的下调，导致线粒体功能障碍，进而导致 Endo G 和 AIF 的释放，从而引发细胞凋亡。
