【摘要】 目的：转录因子STAT5在慢性粒细胞白血病(chronic myeloid leukemia,CML)中组成性激活,并与CML细胞恶性表型密切相关,本研究用cDNA芯片检测方法探讨诱骗核苷酸(decoy ODNs)抑制STAT5对白血病K562细胞株凋亡相关基因的影响。方法：分别提取decoy ODNs处理前后的K562细胞总RNA,逆转录合成Cy3、Cy5标记的cDNA探针,与人14K基因表达谱cDNA芯片(V2.0)杂交,扫描获得数据后,用Genespring软件分析对照组和实验组的差异表达基因,半定量RT- PCR验证cDNA芯片结果。结果：2张cDNA芯片检测重复性良好(R=0.9799)。在13824个标记的基因中,检出413个上调基因,其中包括：TIAF1、GAB1、GYPA、DAPK3、TNFRSF1B、IRF1和PML等在内的18个凋亡相关基因；在检出的332个下调基因中,发现PIM1、CCND2、CCND1、MYC和BCL2L1等在内的11个凋亡相关基因；部分基因经半定量RT-PCR证实与cDNA芯片结果一致。结论：Decoy ODNs抑制STAT5信号通路后可引起K562细胞多种凋亡相关基因的变化,本实验为进一步研究STAT5信号通路所调控的凋亡相关基因提供了依据。更多还原

【Abstract】 Objective:Constitutive activation of signal tranducers and activators of transcription 5(STAT5)has close rela- tionship with the malignant behavior of chronic myeloid leukemia(CML)cells.The present study aimed to map the expression profile of apoptosis-related genes after inhibition of STAT5 by decoy oligodeoxynucleotides(ODNs)in leukemia K562 ceils by u- sing cDNA microarray technique.Methods:Total RNA was extracted from K562 cells before and after decoy ODNs treatment. Two eDNA probes,labeled with Cy3 or Cy5 fluorescent dye,were synthesized via reverse transcription and hybridized with hu- man 14K eDNA microarry.Differential gene expression profiles of decoy ODNs group and control group were analyzed by Gene- spring software.Semiquantitative RT-PCR was used to confirm the result of selected genes from microarray analysis.Results: The cDNA microarray data of the two chips showed good repeatability(R=0.9799).Out of the labeled 13,824 genes,413 genes were up-regulated including 18 apoptosis-related genes such as TIAFI,GAB1,GYPA,DAPK3,TNFRSF1 B,IRF1,and PML,etc. From 332 down-regulated genes,11 genes(PIM1,CCND2,CCND1,MYC,BCL2L1,etc)were related with apoptosis.The mi- croarray results of some selected genes were verified by semiquantitative RT-PCR.Conclusion:Inhibition of STATS signal path- way by decoy ODNs causes changes of many apoptosis-related genes in K562 cells.These results provide evidence for further an- alyzing apoptosis-related genes regulated by STAT5.更多还原