Novel Multiple Apoptotic Mechanism of Shikonin in Human Glioma Cells
Novel Multiple Apoptotic Mechanism of Shikonin in Human Glioma Cells
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DOI：
10.1245/s10434-012-2324-4
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发表时间：
2012-09-01
影响因子：
3.7
通讯作者：
Yang, Jen-Tsung
中科院分区：
文献类型：
--
作者：
Chen, Ching-Hsein;Lin, Miao-Ling;Yang, Jen-Tsung
紫草素是紫草根中主要的萘醌类化合物。我们前期的研究表明紫草素对人肝癌细胞具有一定的抗肿瘤活性。但紫草素对人脑胶质瘤细胞的抗肿瘤作用机制目前尚不清楚。本研究采用流式细胞仪检测紫草素诱导U87 MG、Hs 683和M059 K细胞凋亡、细胞周期、活性氧（ROS）产生、谷胱甘肽（GSH）耗竭和线粒体跨膜电位的改变。凋亡相关蛋白、过氧化氢酶和超氧化物歧化酶-1（SOD-1）通过Western blot检测。应用N-乙酰半胱氨酸（NAC）、pifithrin-alpha（PFT-alpha）和环孢菌素A（cyclosporin A）等药物，研究紫草素诱导U87 MG胶质瘤细胞凋亡的分子机制，结果表明紫草素诱导U87 MG细胞产生ROS、耗竭GSH、破坏线粒体跨膜电位、上调p53和裂解PARP [poly（ADP-ribose）polymerase]。此外，紫草素引起过氧化氢酶下调和SOD-1上调，以及Bcl-2和Bax的表达增加减少。用NAC、PFT-α或环孢菌素A预处理引起紫草素诱导的细胞凋亡的恢复。紫草素诱导的ROS产生和GSH耗竭触发线粒体跨膜电位的破坏。这些研究首次表明紫草素通过多种途径诱导人脑胶质瘤细胞凋亡。因此紫草素有可能作为一种潜在的抗脑胶质瘤的化疗药物。
Shikonin is the main naphthoquinone compound of the root of Lithospermum erythrorhizon. Our previous study demonstrated that shikonin possesses anticancer activity in human hepatoma cells. However, the anticancer mechanism of shikonin in human glioma cells is unclear at present. In the present study, we demonstrated that shikonin induces apoptosis in three human glioma cell lines: U87MG, Hs683, and M059K cells.Cell cycle, generation of reactive oxygen species (ROS), depletion of glutathione (GSH), and disruption of mitochondrial transmembrane potential in shikonin-treated cells were determined by flow cytometry. Apoptosis-related proteins, catalase, and superoxide dismutase-1 (SOD-1) were determined by Western blot testing. N-acetylcysteine (NAC), pifithrin-alpha (PFT-alpha), or cyclosporin A were applied to evaluate the molecular mechanism of shikonin in apoptosis.Shikonin induces the generation of ROS, depletion of GSH, disruption of mitochondrial transmembrane potential, upregulation of p53, and cleavage of PARP [poly(ADP-ribose) polymerase] in U87MG glioma cells. Moreover, shikonin causes catalase downregulation and SOD-1 upregulation as well as decreased Bcl-2 and increased Bax expression. Pretreatment with NAC, PFT-alpha, or cyclosporin A causes the recovery of shikonin-induced apoptosis. The ROS generation and GSH depletion induced by shikonin trigger mitochondrial transmembrane potential disruption. ROS production was partially dependent on the upregulation of p53 upon shikonin treatment.These studies are the first to show that shikonin-induced apoptosis occurs through multiple pathways in human glioma cells. We conclude that shikonin may be used as a potential chemotherapeutic agent against human glioma.