三氧化二砷对急性早幼粒细胞白血病细胞凋亡相关基因表达的影响

Effect of Arsenic Trioxide on the Expression of Apoptosis-Related Genes in NB4 cells

本研究采用微矩阵基因芯片技术探讨三氧化二砷(As2O3)对NB4细胞凋亡的相关基因表达谱的影响。As2O3作用NB4细胞48小时,用改进的TRIZOL试剂一步法抽提As2O3作用前后的NB4细胞总RNA;经逆转录-聚合酶链反应后用Cy3标记的脱氧三磷酸尿苷(Cy32dUTP)和Cy52dUTP两种不同的荧光染料,将As2O3作用前后的NB4细胞mRNA分别标记成两种DNA探针,并与载有一组凋亡相关基因的表达谱芯片进行杂交,扫描分析筛选As2O3作用前后NB4细胞表达差异的凋亡基因;采用逆转录实时定量聚合酶链反应(RQ-PCR)检测p21,survivin,cdc2及Wee1 Hu等方法检测细胞凋亡。结果表明:从As2O3作用后NB4细胞中筛选出表达差异的凋亡相关基因共18条,包括p21,survivin,cdc2及Wee1 Hu等,表达上调的有6条,表达下调有12条;p21、survivin、cdc2及Wee1 Hu可能与As2O3诱导NB-4细胞的分化和(或)凋亡有关。结论:As2O3介导的NB4细胞凋亡的发生机制中、p21、sur-vivin、cdc2及Wee1可能发挥着重要作用。

The aim of this study was to investigate the gene expression profiles of acute promyelocytic leukemia （APL） cell line NB4 treated with arsenic trioxide （As2O3 ） by using cDNA microarray, cDNA probes were prepared through reverse transcription from mRNA of NB4 cells treated with or without arsenic trioxide. The probes were labeled with Cy3 and Cy5 fluorescence dyes individually, hybridized with cDNA microarray representing 201 different human genes, and their fluorescent intensities were scanned. The genes were screened through the analysis of the difference in the gene expression profile. The results showed that after the treatment of arsenic trioxide （2 μmol/L）, 6 genes were up-regulated, and 12 genes related to apoptosis and signal transduction were down-regulated. The p21, survivin, cdc2 and WeelHu genes may be related to the differentiation and/or apoptosis of NB4 cells induced by As：O3. It is concluded that p21, survivin, cdc2 and Weel Hu may play an important role in the mechanism underling arsenic trioxide-mediated NB4 cell apoptosis.