cDNA芯片检测STAT5诱骗核苷酸对K562细胞凋亡相关基因的影响  

Effect of STAT5 decoy oligodeoxynucleotides on expression profile of apoptosis-related genes in K562 cells by cDNA microarray

作　　者：曾建明[1] 冯文莉[1] 王小中[1] 史梅[1] 涂植光[2] 黄宗干[3] 

机构地区：[1]重庆医科大学临床血液学教研室,重庆400016 [2]重庆医科大学临床生物化学教研室,重庆400016 [3]重庆医科大学临床学院血液科,重庆400016

出　　处：《肿瘤》2006年第8期738-742,共5页Tumor

基　　金：国家自然科学基金(编号:30270574)

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。Objective：Constitutive activation of signal tranducers and activators of transcription 5 （STATS） has close relationship 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 STATS by decoy oligodeoxynucleotides （ODNs） in leukemia K562 cells by using eDNA 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 human 14K cDNA microarry. Differential gene expression profiles of decoy ODNs group and control group were analyzed by Genespring software. Semiquantitative RT-PCR was used to confirm the result of selected genes from microarray analysis. Results： The eDNA 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 TIAF1, GAB1, GYPA, DAPK3, TNFRSF113, IRF1, and PML, etc. From 332 down regulated genes,ll genes （PIM1,CCND2,CCND1,MYC, BCL2L1,etc） were related with apoptosis. The microarray results of some selected genes were verified by semiquantitative RT PCR. Conclusion：Inhibition of STATS signal pathway by decoy ODNs causes changes of many apoptosis-related genes in K562 cells. These results provide evidence for further analyzing apoptosis-related genes regulated by STATS.

关 键 词：白血病 髓样 慢性 信号转导和转录活化因子 诱骗脱氧寡核苷酸 细胞凋亡 寡核苷类排列序列分析 K562细胞 

分 类 号：R577.1[医药卫生—消化系统] R733.72[医药卫生—内科学]