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  

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

作　　者：陈将华 郑维威[1] 姜旭东[1] 陆晓雅[1] 徐荣臻[1] 

机构地区：[1]浙江大学医学院附属第二医院血液科,浙江杭州310009

出　　处：《南方医科大学学报》2015年第5期702-706,共5页Journal of Southern Medical University

基　　金：国家自然科学基金(81270601;81470306)~~

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Origin Pro8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m RNA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m RNA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 m RNA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 m RNA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。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.

关 键 词：雷公藤甲素 急性T淋巴细胞白血病Jurkat细胞 凋亡 HERV-K Np9基因 

分 类 号：R96[医药卫生—药理学]