南方医科大学学报

南方医科大学学报 ›› 2015, Vol. 35 ›› Issue (05): 702-.

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雷公藤甲素通过抑制逆转录病毒HERV-K Np9基因转录诱导人急性T淋巴细胞白血病Jurkat细胞凋亡

陈将华，郑维威，姜旭东，陆晓雅，徐荣臻   

  • 出版日期:2015-05-20 发布日期:2015-05-20

Triptolide induces apoptosis of human acute T lymphocytic leukemia Jurkat cells via
inhibiting transcription of human endogenous retrovirus HERV-K Np9 gene

  • Online:2015-05-20 Published:2015-05-20

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摘要： 目的探讨雷公藤甲素诱导人急性T淋巴细胞白血病Jurkat细胞凋亡分子机制。方法MTT检测雷公藤甲素对Jurkat细
胞的增殖抑制作用，然后用OriginPro8计算出IC50。按照0、2、4、8、16 nmol/L浓度雷公藤甲素处理Jurkat细胞48 h，然后用流式
细胞仪检测细胞凋亡变化。用半定量RT-PCR法检测加药处理后各组Np9基因mRNA的表达水平变化并用Kodak 1D 3.6软件
对条带进行定量分析。采用统计软件分析Np9转录抑制与细胞凋亡的相关性。Western Blotting检测Np9下游信号分子c-myc，
β-catenin，ERK，AKT和Notch1蛋白的变化。结果雷公藤甲素呈剂量依赖性抑制Jurkat细胞的增殖，其IC50为12.7 nmol/L。雷公
藤甲素呈剂量依赖诱导Jurkat细胞凋亡。进一步实验研究结果显示，雷公藤甲素呈剂量依赖方式抑制Jurkat细胞中Np9基因的
mRNA的转录水平。经统计分析发现Np9转录抑制与细胞凋亡之间具有显著的相关性（R2=0.907）。Western Blotting方法结果
发现雷公藤甲素在抑制Np9 mRNA转录同时伴有其下游信号分子c-myc，β-catenin，ERK，AKT和Notch1蛋白表达水平降低。
结论下调HERV-K Np9 mRNA及其下游信号分子c-myc，β-catenin，ERK，AKT和Notch1蛋白水平是雷公藤甲素诱导人急性T
淋巴细胞白血病Jurkat细胞凋亡的重要分子机制之一。

Abstract: Objective To investigate the molecular mechanisms by which triptolide induces apoptosis of human acute T
lymphocytic leukemia Jurkat cells. Methods MTT assay was employed to detect the proliferation inhibition of Jurkat cells by
triptolide, and the IC50 was calculated by OriginPro8. Flow cytometry was used to analyze apoptosis of Jurkat cells. Np9
mRNA levels were detected by RT-PCR and analyzed quantitatively by Kodak 1D 3.6 software. Correlation between the
inhibition of Np9 transcription and the cell apoptosis was analyzed by SPSS 19.0.Western blotting was employed to determine
Np9 downstream signaling molecules c-myc, β-catenin, ERK, AKT and Notch1 protein level in Jurkat cells after exposure to
different concentrations of triptolide for 48h. Results Triptolide treatment resulted in dose-dependent inhibition of Jurkat cells
proliferation and its IC50 was 12.7nmol/L. Triptolide induced apoptosis of Jurkat cells in dose- dependent manner. Furthermore,
triptolide inhibited Np9 mRNA transcription level in Jurakt cells in a dose-dependent manner. There was a correlation
between the triptolide-mediated the apoptosis and the inhibition of Np9 transcription of Jurkat cells (R2=0.907). Western
blotting results displayed that triptolide inhibited transcription levels of Np9 mRNA with a concomitant decrease of its
downstream signaling molecules c-myc, β-catenin, ERK, AKT and Notch1 at protein levels. Conclusion Inhibition of HERV-K
Np9 mRNA and its downstream signaling molecules c-myc, β-catenin, ERK, Akt and Notch1 protein might be one of
important molecular mechanisms by which triptolide induces apoptosis of human acute T lymphocytic leukemia Jurkat cells.