| 熊春雷,姚青林,冯 品,王晓光,张静宜.大蒜素诱导人白血病HL-60细胞程序性坏死及分子机制的研究[J].,2017,17(35):6833-6838 |
| 大蒜素诱导人白血病HL-60细胞程序性坏死及分子机制的研究 |
| Study on Allicin-induced Necroptosis and Related Molecular Mechanism in Human Leukemia HL-60 Cells |
| 投稿时间：2017-06-28 修订日期：2017-07-22 |
| DOI：10.13241/j.cnki.pmb.2017.35.008 |
| 中文关键词: 大蒜素 白血病 程序性坏死 RIP1 JNK HL-60细胞 |
| 英文关键词: Allicin leukemia Necroptosis RIP1 JNK HL-60 cells |
| 基金项目:陕西省自然科学基金项目（2016JM3750） |
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| 中文摘要: |
| 摘要 目的：研究大蒜素对人白血病HL-60细胞的抗肿瘤作用，检测程序性坏死明确大蒜素处理后HL-60细胞的死亡方式，并以JNK、RIP1和RIP3为靶点探讨可能的分子机制。方法：不同浓度大蒜素处理HL-60细胞后，在不同时间点采用MTT法检测细胞活力，使用非选择性caspase抑制剂z-vad-fmk研究细胞凋亡在其中的作用；50 μM大蒜素处理HL-60细胞后，检测LDH释放量和PI染色阳性细胞比例反映细胞程序性坏死程度，采用免疫印迹法检测各时间点RIP1、RIP3表达和JNK磷酸化程度；使用JNK特异性抑制剂SP600125处理HL-60细胞，采用免疫共沉淀法检测RIP1与RIP3相互作用，并通过检测细胞活力、LDH释放量和PI染色阳性细胞比例研究JNK在大蒜素抗白血病活性中的作用。结果：大蒜素可显著抑制人白血病HL-60细胞增殖，这种作用并不完全依赖于诱导细胞凋亡；50 μM大蒜素可诱导HL-60细胞发生程序性坏死，这种作用可被程序性坏死的特异性抑制剂necrostatin-1逆转；50 μM大蒜素可显著增加HL-60细胞RIP1的表达和JNK的磷酸化水平，而对RIP3的表达无明显影响；50 μM大蒜素可显著增加RIP1与RIP3的相互作用，使用JNK特异性抑制剂SP600125可逆转大蒜素的抗白血病作用。结论：一定剂量的大蒜素可通过激活JNK增加RIP1与RIP3的相互作用，诱导人白血病HL-60细胞发生程序性坏死，进而发挥抗白血病作用。 |
| 英文摘要: |
| ABSTRACT Objective: The aim of this study was to investigate the anti-cancer effect of allicin in human leukemia HL-60 cells. We assayed necroptosis to determine the cell death type induced by allicin in HL-60 cells, and investigated the underlying molecular mecha- nisms with focus on JNK, RIP1 and RIP3 proteins. Methods: After treatment with allicin at different concentrations, the cell viability in HL-60 cells was measured by MTT method at different time points. The pan caspase inhibitor z-vad-fmk was used to investigate the in- volvement of anti-apoptotic mechanism. After treatment with 50 μM allicin, the LDH release and PI-positive cells was measured to de- tect necroptosis in HL-60 cells, and western blot was performed to detect the expression of RIP1 and RIP3, and the phosphorylation of JNK. HL-60 cells were treated with the selective JNK inhibitor SP600125, and co-immunoprecipitation (Co-IP) was performed to detect the interaction of RIP1 with RIP3. The cell viability, LDH release and PI-positive cells was measured after allicin and SP600125 treat- ment to investigate the involvement of JNK in allicin-induced anti-cancer effects in HL-60 cells. Results: Allicin exerted anti-cancer ef- fects in human leukemia HL-60 cells, and some apoptosis-independent mechanisms were also involved in. Allicin at 50 μM induced necroptosis in HL-60 cells, which could be reversed by the necroptosis inhibitor necrostatin-1. Allicin at 50 μM significantly increased the expression of RIP1 and the phosphorylation of JNK, but had no effect on RIP3 expression. Allicin at 50 μM significantly enhanced the interaction of RIP1 with RIP3, and the selective JNK inhibitor SP600125 could reverse the anti-cancer effects of allicin in HL-60 cells. Conclusion: Allicin induced necroptosis through JNK-dependent enhancement of RIP1-RIP3 interaction, which contributed to the anti-cancer effects of allicin in human leukemia HL-60 cells. |
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