隐丹参酮通过激活依赖于动力相关蛋白 1 的线粒体碎片化来抑制骨肉瘤。
Activation of dynamin-related protein 1 - dependent mitochondria fragmentation and suppression of osteosarcoma by cryptotanshinone.
机构信息
Research Cancer Center for Traditional Chinese Medicine, Department of Medical Research, China Medical University Hospital, Taichung, Taiwan.
Department of Chinese Medicine, China Medical University Hospital, No. 2, Yude Rd, North District, Taichung, 40447, Taiwan.
出版信息
J Exp Clin Cancer Res. 2019 Jan 28;38(1):42. doi: 10.1186/s13046-018-1008-8.
BACKGROUND
Discovering how to regulate mitochondrial function to reduce cancer growth holds great potential for future cancer therapy development. Here we explore the effects of cryptotanshinone (CPT), a natural product derived from Salvia miltiorrhiza, on mitochondria of osteosarcoma (OS) both in vitro and in vivo, and further elucidate the underlying molecular mechanisms.
METHODS
Cytotoxicity in the CPT treated OS cells was analyzed by flow cytometry, CCK8, TUNEL assay and colony formation assays. Flow cytometric analysis was performed to evaluate the effect of CPT on cell cycle of OS cells. Mitochondrial morphology was examined by staining with the mitochondrial membrane potential -sensitive fluorochrome, MitoTracker Red (CMXRos). Immunoblotting, confocal-immunofluorescence staining, co-immunoprecipitation were used to examine the expression and interaction between CPT-mediated Drp1 and Bax. Finally, the synergistic effect of CPT on OS cells was validated using a mouse xenograft tumor model.
RESULTS
In this study, we found CPT treatment induced S-phase arrest, apoptosis, and mitochondrial fragmentation in OS cells. CPT also effectively activated caspase-dependent apoptosis, which could be blocked by pan-caspase inhibitor Z-VAD-FMK. Moreover, we herein provide evidence that treatment with CPT resulted in mitochondrial fragmentation, which is mediated by dynamin-related protein 1 (Drp1), a key mediator of mitochondrial fission. Pursuing this observation, downregulation of Drp1 via silencing RNA could abrogate the induction of apoptosis and mitochondrial fragmentation induced by CPT. Finally, we demonstrate that CPT induced Drp1, which interacted directly with Bcl-2-associated X protein (Bax), which contributed to driving Bax translocation from the cytosol to the mitochondria.
CONCLUSIONS
Our findings offer insight into the crosstalk between mitochondrial fragmentation and inhibition of osteosarcoma cell growth in response to CPT.
背景
探索如何调节线粒体功能以减少癌症生长，这对于未来癌症治疗的发展具有巨大潜力。在这里，我们研究了来源于丹参的天然产物隐丹参酮（CPT）对骨肉瘤（OS）细胞线粒体的体内外作用，并进一步阐明了潜在的分子机制。
方法
通过流式细胞术、CCK8、TUNEL 测定和集落形成测定分析 CPT 处理的 OS 细胞中的细胞毒性。通过流式细胞术分析评估 CPT 对 OS 细胞细胞周期的影响。用线粒体膜电位敏感荧光染料 MitoTracker Red（CMXRos）染色来检测线粒体形态。用免疫印迹、共聚焦免疫荧光染色、共免疫沉淀来检测 CPT 介导的 Drp1 和 Bax 之间的表达和相互作用。最后，使用小鼠异种移植肿瘤模型验证 CPT 对 OS 细胞的协同作用。
结果
在这项研究中，我们发现 CPT 处理诱导 OS 细胞的 S 期阻滞、凋亡和线粒体碎片化。CPT 还能有效地激活依赖半胱氨酸的天冬氨酸蛋白水解酶（caspase）依赖性凋亡，这可被广谱半胱氨酸天冬氨酸蛋白酶抑制剂 Z-VAD-FMK 阻断。此外，我们在此提供证据表明，CPT 处理导致线粒体碎片化，这是由线粒体裂变的关键介质——动力相关蛋白 1（Drp1）介导的。通过沉默 RNA 下调 Drp1 可以阻断 CPT 诱导的凋亡和线粒体碎片化的诱导。最后，我们证明 CPT 诱导 Drp1 与 Bcl-2 相关 X 蛋白（Bax）直接相互作用，导致 Bax 从细胞质易位到线粒体。
结论
我们的研究结果深入了解了线粒体碎片化与 CPT 抑制骨肉瘤细胞生长之间的相互作用。
Zotero 插件