2-甲氧基雌二醇及三氧化二砷对骨髓瘤细胞凋亡相关基因表达谱影响的研究

Investigation of the effect of 2-methoxyestradiol and arsenic trioxide on the apoptosis-associated gene expression profile of myeloma cells

目的 探讨2 甲氧基雌二醇(2ME2)及三氧化二砷(As2O3 )对多发性骨髓瘤(MM)细胞系CZ 1细胞凋亡相关基因表达的影响。方法 应用TRIzol一步法抽提经凋亡剂量的2ME2及As2O3处理后CZ 1细胞总RNA,经逆转录合成Bio 16 dUTP标记的cDNA探针,与微矩阵排列的含有96种凋亡相关基因的表达谱芯片杂交,利用GEArrayAnalyzer软件分析2ME2及As2O3 处理对CZ 1细胞凋亡相关基因表达的影响及差异,并进一步用RT PCR法证实。结果 As2O3 诱导了52条(占芯片凋亡相关基因总数的54. 2% )凋亡相关基因表达发生改变,其中42条(占表达改变的凋亡相关基因总数的80. 8% )基因表达上调及10条( 19. 2% )基因表达下调;上调的基因主要涉及caspases家族、P53及ATM通路、死亡结构域家族、TNF受体家族及CIDE家族等。2ME2诱导了42条(占芯片凋亡相关基因总数的43. 8% )凋亡相关基因表达改变,其中32条(占表达改变的凋亡相关基因总数的76. 2% )基因表达下调及10条(23. 8% )基因表达上调;下调基因主要涉及bcl 2家族、IAP家族、TRAF家族、TNF配体家族及CARD家族等。结论 2ME2及As2O3 分别通过不同的凋亡通路诱导CZ 1细胞凋亡。As2O3主要诱导一些促凋亡基因表达上调;而2ME2则主要下调了一些抗凋亡基因表达。

Objective To explore the effect of 2-methoxyestradiol (2ME2) and arsenic trioxide(As 2O 3) on the apoptosis related gene expression in multiple myeloma cell line CZ-1. Methods Total RNA was isolated from CZ-1 cells which had been treated with 2ME2 and As 2O 3 at an apoptosis-inducing concentration and reverse-transcribed into a cDNA probe labeled with Bio-16-dUTP, and then hybridized it with a microarray containing up to 96 key genes involved in apoptosis. The gene expression profile of the 2ME2 and As 2O 3 treated CZ-1 cells were analyzed with GEArray Analyzer software. The microarray results were comfirmed by RT-PCR. Results As 2O 3 treatment caused the alteration in the expression of 52 genes (54.2% of total genes on microarray). Among them, 42 (80.8%) were upregulated and 10 (19.2%) were downregulated. The upregulated genes were mainly involved in caspases family, P53 and ATM pathway, death effector domain family,TNF receptor family and CIDE family. 2ME2 treatment resulted in the alteration of 42 genes (43.8% of the total genes on microarray). Of them, 32 (76.2%) were downregulated and 10 ( 23.8% ) were upregulated. The downregulated genes mainly belonged to bcl-2 family, inhibitor of apoptosis protein family (IAP), TRAF family,TNF ligand family, and CARD family. Conclusion 2ME2 and As 2O 3 induce the CZ-1 cells apoptosis by different pathways. As 2O 3 mainly induces upregulation of proapoptotic genes, and 2ME2 downregulation of anti-apoptotic genes expression.