人参皂甙对K562细胞作用的基因表达谱研究

Gene expression profiles of k562 cells treated with total saponin of Panax ginseng

目的运用基因芯片技术研究人参总皂甙(TSPG)作用后K562细胞基因表达谱的变化。方法200 μg/mlTSPG作用于K562细胞3d后,提取总RNA,合成cRNA并分别用cy3和cy5标记,与Agilent Human 1B寡核苷酸基因芯片杂交,研究基因表达谱的变化。结果TSPG刺激K562细胞后共有362个基因表达发生变化,与对照组K562细胞比较,表达上调的基因有20个,主要有代谢相关基因,信号转导相关基因,细胞受体相关基因等;表达下调的有342个,包括免疫防御相关基因,DNA结合与转录因子,代谢相关基因,细胞周期相关基因等。RT-PCR技术验证了FOSL1、E2F2、CCNE2和ODZ1四个基因表达的变化。结论TSPG刺激K562细胞后,影响了细胞内一系列基因的表达。这些基因可能与TSPG的抗肿瘤机制有关。

Objective To investigate the effect of the total saponin of Panax ginseng （TSPG） on gene expression profile of K562 cells using microarray technique. Methods The total RNA were extracted and purified from K562 cells treated by 200 μg/ml TSPG for 3 days, and untreated K562 cells cultured in parallel served as the control, cRNAs were synthesized and labeled with Cy3 and Cy5 respectively. The labeled eRNA fragments were hybridized with Agilent human 1B 60 mer oligonucleotide microarray, which was then scanned to reveal the changes of gene expression profile in relation to TSPG treatment. Results Totally 362 differentially expressed genes were identified in TSPG-treated K562 cells, including 20 up-regulated ones （consisting of metabolism-associated genes, signal transducfion-associated genes and cell receptor-associated genes etc） and 342 down-regulated ones （consisting of immunity and defense-associated genes, DNA-binding and transcription genes, metabolism-associated genes and cell cycle-associated genes etc）. Changes in expressions ofFOSL1, E2F2, CCNE2 and ODZI were confirmed by semi-quantitative RT-PCR. Conclusions TSPG may induce changes in the gene expression profile in k562 cells possibly relevant to the anti-tumor mechanism of TSPG.