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目的:在全基因组范围研究雷公藤内酯醇对Jurkat细胞基因表达的调节作用,筛选雷公藤内酯醇作用的靶分子。方法:将对数生长期的Jurkat细胞分为正常对照及雷公藤内酯醇处理组,后者同时加入10μg/L的雷公藤内酯醇并在37℃温育2h,从两组细胞中分别分离总RNA,并以此为模板逆转录成荧光标记的cDNA探针,将上述探针分别与含13872个人类基因/Est的DNA芯片进行杂交,所获DNA芯片进行杂交影像经DNA芯片数据分析软件GeneSpring进行分析,得到基因表达图谱。结果:雷公藤内酯醇显著抑制117个基因/Est在Jurkat细胞中的表达,在雷公藤内酯醇调节的转录产物中30％为Est或末知功能基因,13％为转录因子,9％为信号转导途径的调节分子,9％为DNA结合蛋白,十分引人注目的是MAPK5和PⅠ-3K的表达被下调了100 倍以上,除此之外,雷公藤内酯醇还能抑制参与脂类转移及代谢的基因的表达。结论:高密度DNA芯片技术是一种有效的药物作用靶分子的筛选手段,雷公藤内酯醇有可能通过其对MAPK和PⅠ-3K的抑制作用发挥免疫抑制和抗肿瘤作用。
OBJECTIVE: To study the regulatory effect of triptolide on the gene expression of Jurkat cells in the genome-wide range and to screen the target molecules of triptolide. Methods: Jurkat cells in logarithmic growth phase were divided into normal control group and triptolide treated group. At the same time, 10 μg / L triptolide was added to the Jurkat cells and incubated at 37 ℃ for 2 h. RNA as a template reverse transcribed into fluorescently labeled cDNA probe, respectively, the probe with 13872 human genes / Est DNA chip hybridization, the obtained DNA chip hybrid image DNA chip data analysis software GeneSpring For analysis, get the gene expression profile. Results: Triptolide significantly inhibited the expression of 117 genes / Est in Jurkat cells, 30% of which were Est or terminal function genes in triptolide-regulated transcripts, 13% were transcription factors and 9% were signal In addition, triptolide also inhibits the expression of MAPK5 and PI-3K, which is down-regulated more than 100-fold. In addition, triptolide also inhibits the expression of MAPK5 and PI- And metabolic gene expression. Conclusion: High-density DNA chip technology is an effective screening method for target molecules of drugs. Triptolide may exert immunosuppressive and anti-tumor effects through inhibition of MAPK and PⅠ-3K.