| Status | 已發表Published |
| An NQO1 dependent ROS and RIP1/RIP3 mediated necroptosis induced in glioma cancer cells by a natural product, 2-Methoxy-6-acetyl-7-methyljuglone (MAM) | |
Yu, J. ; Li, Z.W.; Hou, Y.; Yuan, R.Y.K.; Lu, J. ; Chen, X. P.
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| 2017-11-01 | |
| Source Publication | Collection The 6th Guangzhou International Symposium on Oncology
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| Abstract | ckground: Glioblastomas (GBM) are the most malignant brain tumors in humans and have a very poor prognosis. New therapeutics are urgently needed. 2-methoxy-6-acetyl-7-methyljuglone (MAM) was reported to induce necroptosis in colon cancer cells, but no report in glioma cancer cells. Thus, it is needed to clarify whether MAM could cause necroptosis in glioma cancer cells and investigated its underlying mechanisms. Methods: MAM and human U87 glioma cancer cell line and U251 human glioma cancer cell line were used in this study. The cellular viability was assayed by MTT. Flow cytometry with Annexin V and 7AAD double staining was used to analyze cell death type. Morphological alterations were evaluated by fluorescence microscopy with Hoechst 33342 and PI double staining. The level of reactive oxygen species (ROS) and calcium level was analyzed by using redox-sensitive dye DCFH2-DA and calcium dye fluo-3AM. The expression level of necroptosis associated proteins was analyzed by western blotting. Results: MAM induced cell death in U87 and U251 glioma cancer cells without caspase activation. The cell death induced by MAM was attenuated by the pharmacological or genetic blockage of necroptosis signaling, including RIP1 inhibitor necrostatin-1s (Nec-1s) and siRNA-mediated gene silencing of RIP1 and RIP3, but was unaffected by caspase inhibitor z-vad-fmk or necrosis inhibitor 2-(1H-Indol-3-yl)-3-pentylamino-maleimide (IM54). MAM treated U87 and U251 glioma cancer cells induced RIP1/RIP3 complex formation, ROS level increased, ATP concentration decreased and loss of plasma membrane integrity, further confirmed this process was necroptosis. The essential role of ROS was confirmed by the protective effect of ROS scavenger NAC. Moreover, MAM induced necroptosis both triggered by RIP1/RIP3 complex and ROS generation. MAM induced necroptosis through cytosolic calcium (Ca2+) accumulation and sustained c-Jun N-terminal kinase (JNK) activation. Both calcium chelator BAPTA-AM and JNK inhibitor SP600125 could attenuate cell death. Finally, MAM induced necroptosis was inhibited by dicoumarol (a NQO1 inhibitor). Dicoumarol exposed glioma cancer cells were resistant to RIP1/RIP3 complex formation and ROS generaion. Conclusions: Take together, our results demonstrated that MAM induced necroptosis through the NQO1-dependent ROS and RIP1/RIP3 pathway. Our study also provided new insights into the molecular regulation of necroptosis in human glioma cancer cells and a promising approach for GBM treatment. |
| Keyword | MAM necroptosis NQO1 ROS RIP1/RIP3 |
| Language | 英語English |
| The Source to Article | PB_Publication |
| PUB ID | 39357 |
| Document Type | Conference paper |
| Collection | Institute of Chinese Medical Sciences |
| Corresponding Author | Chen, X. P. |
| Recommended Citation GB/T 7714 | Yu, J.,Li, Z.W.,Hou, Y.,et al. An NQO1 dependent ROS and RIP1/RIP3 mediated necroptosis induced in glioma cancer cells by a natural product, 2-Methoxy-6-acetyl-7-methyljuglone (MAM)[C], 2017. |
| APA | Yu, J.., Li, Z.W.., Hou, Y.., Yuan, R.Y.K.., Lu, J.., & Chen, X. P. (2017). An NQO1 dependent ROS and RIP1/RIP3 mediated necroptosis induced in glioma cancer cells by a natural product, 2-Methoxy-6-acetyl-7-methyljuglone (MAM). Collection The 6th Guangzhou International Symposium on Oncology. |
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