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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 investigate the effects of 2methoxyestradiol (2ME2) and arsenic trioxide (As2O3) on the expression of apoptosis related genes in multiple myeloma (MM) cell line CZ1. Methods The total RNA of CZ 1 cells treated with 2ME2 and As 2 O 3 at the apoptotic doses was extracted by one-step TRIzol method. The cDNA probes containing Bio 16 dUTP labeled with cDNA were reverse transcribed, and the expression of 96 apoptosis-related genes Spectrum chip hybridization, the use of GEArrayAnalyzer software analysis 2ME2 and As2O3 treatment of CZ1 cells apoptosis-related gene expression and differences, and further confirmed by RT PCR. Results As2O3 induced 52 apoptosis-related genes (accounting for 54.2% of the total number of apoptosis-related genes), and 42 of them (80.8% of the total number of apoptosis-related genes) were up-regulated And 10 (19.2%) were down-regulated. The up-regulated genes mainly involved caspases family, P53 and ATM pathways, death domain family, TNF receptor family and CIDE family. 2ME2 induced the expression of 42 apoptosis-related genes (accounting for 43.8% of the total number of apoptotic-related genes), of which 32 (accounting for 76.2% of the total number of apoptotic-related genes with altered expression) were down-regulated and 10 (23.8%) gene expression was up-regulated; down-regulated genes mainly involved bcl 2 family, IAP family, TRAF family, TNF ligand family and CARD family. Conclusion 2ME2 and As2O3 induce CZ 1 cell apoptosis through different apoptotic pathways respectively. As2O3 mainly induces the up-regulation of some pro-apoptotic genes, while 2ME2 mainly down-regulates some anti-apoptotic gene expression.