| 郑璟瑶.青蒿素抑制CD133+HepG2细胞对γδ T细胞的耐受性及其机制研究[J].浙江中西医结合杂志,2017,27(12): |
| 青蒿素抑制CD133+HepG2细胞对γδ T细胞的耐受性及其机制研究 |
| The effect and mechanism of artemisinin on reducing the tolerance of CD133+ HepG2 to γδ T cells |
| 投稿时间：2017-06-04 修订日期：2017-06-04 |
| DOI： |
| 中文关键词: 青蒿素 MCL-1 γδ T细胞 CD133 HepG2 |
| 英文关键词:artemisinin MCL-1 γδ T cells CD133 HepG2 |
| 基金项目: |
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| 中文摘要: |
| 目的: 探讨青蒿素是否能抑制CD133+肝癌细胞对γδ T细胞的耐受性及研究其机制。方法：LDH释放实验检测γδ T细胞及青蒿素共培养对CD133+及CD133- HepG2的杀伤活性；western blot实验检测γδ T细胞及青蒿素共培养对CD133+ HepG2细胞MCL-1的表达水平、caspase-9、caspase-3的活化水平和细胞色素c的释放水平的影响；流式细胞实验检测γδ T细胞及青蒿素共培养对CD133+ HepG2细胞凋亡和线粒体膜电位的影响。结果：LDH释放实验结果显示在相同数量γδ T细胞处理下,CD133+ HepG2细胞的LDH释放率显著低于CD133- HepG2细胞,表明CD133+ HepG2细胞对γδ T细胞治疗存在耐受性；同时,γδ T细胞+青蒿素组CD133+ HepG2的LDH释放率(55.3±6.1)显著高于γδ T细胞组(18.7±2.6, P<0.05)和青蒿素+γδ T细胞+MCL-1质粒组(24.2±2.8, P<0.05)。流式细胞实验结果显示γδ T细胞+青蒿素组CD133+ HepG2的凋亡率(38.2±3.5)显著高于γδ T细胞组(10.5±1.1, P<0.05)和青蒿素+γδ T细胞+MCL-1质粒组(14.3±1.2, P<0.05)。western blot实验结果显示青蒿素处理能显著抑制CD133+ HepG2细胞中MCL-1蛋白的表达水平。另外,流式细胞实验结果显示γδ T细胞+青蒿素组CD133+ HepG2的相对线粒体膜电位(0.21±0.02)显著高于γδ T细胞组(0.78±0.05, P<0.05)和青蒿素+γδ T细胞+MCL-1质粒组(0.71±0.05, P<0.05)。western blot实验结果则显示γδ T细胞+青蒿素组CD133+ HepG2的活化caspase-9 、caspase-3及细胞色素c的释放均显著高于γδ T细胞组和青蒿素+γδ T细胞+MCL-1质粒组。结论：青蒿素可以通过抑制CD133+ 肝癌细胞中MCL-1的表达水平抑制其对γδ T细胞的耐受性。 |
| 英文摘要: |
| AIM: To investigate the effect and mechanism of artemisinin on educing the tolerance of CD133+ HepG2 to γδ T cells. Methods: LDH release assays were performed to evaluate the cell viability of CD133+ HepG2 cells co-cultured with artemisinin and γδ T cells. Western blot analysis was performed to evaluate the expression of MCL-1, activation of caspase-9, caspase-3 and release of cytochrome c in CD133+ HepG2 cells co-cultured with artemisinin and γδ T cells. Flow cytometry analysis was performed to measure the apoptosis and mitochondrial membrane potential in CD133+ HepG2 cells co-cultured with artemisinin and γδ T cells. Results: Results of LDH release assays showed that under the treatment of equal amount of γδ T cells, release rate of LDH in CD133+ HepG2 cells was significantly lower than that in the CD133- HepG2 cells. It indicated that CD133+ HepG2 cells were tolerant to γδ T cells treatment; Meanwhile, the LDH release rate in γδ T cells + artemisinin group (55.3±6.1) was significantly higher than the γδ T cells group (18.7±2.6, P<0.05) and γδ T cells + artemisinin + MCL-1 plasmid group (24.2±2.8, P<0.05). Results of flow cytometry analysis showed that apoptotic rate in γδ T cells + artemisinin group (38.2±3.5) was significantly higher than the γδ T cells group (10.5±1.1, P<0.05) and γδ T cells + artemisinin + MCL-1 plasmid group (14.3±1.2, P<0.05). Results of western blot assays showed that artemisinin downregulated the expression of MCL-1 in CD133+ HepG2 cells. In addition, results of flow cytometry analysis showed that relative mitochondrial membrane potential in γδ T cells + artemisinin group (0.21±0.02) was significantly lower than the γδ T cells group (0.78±0.05, P<0.05) and γδ T cells + artemisinin + MCL-1 plasmid group (0.71±0.05, P<0.05). Results of western blot assays showed that activation of caspase-9, caspase 3, and release of cytochrome c in γδ T cells + artemisinin group was significantly higher than that in the γδ T cells group and γδ T cells + artemisinin + MCL-1 plasmid group. Conclusion: Artemisinin suppressed the tolerance of CD133+ liver cancer cells to γδ T cells through inhibition of MCL-1 expression. |
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