Savli Hakan, Aalto Yan, Nagy Bálint, Knuutila Sakari, Pakkala Seppo
Department of Pathology, Haartman Institute and Helsinki University Central Hospital, Helsinki, Finland.
Br J Haematol. 2002 Sep;118(4):1065-70. doi: 10.1046/j.1365-2141.2002.03734.x.
The alterations in gene expression associated with 1,25(OH)2D3-induced differentiation of HL-60 cells were studied in order to identify potential targets for further investigation of the genetic basis of acute myeloid leukaemia. Atlas human haematology filters, including 406 genes (Clontech), were used to study gene expression in response to 1,25(OH)2D3 (concentration, 5 x 10-8 mol/l) for 24 and 72 h. Compared with untreated cells, expression differences were found in 43 genes. Downregulated genes at both time-points were: IL2RA, CMYC, NPM, DEK, AF4, FLI1, htlf, MNDA, BCR, IKAROS, BPI and NFAT4. Upregulated genes at both time-points were IL1B, CD14 and MCL1. CD55, CD58, IRF2, CREB1, ATF4, RAC1, TIAR, KIAA0053, BAT2, BTK, RCK, EV12B and EDN were downregulated at 24 h, while SPI1, MKK3, BTG1 and IL8 were upregulated. At 72 h the upregulated genes were IL1RA, IL2RG, CXCR4, SCYA1, SCYA3, SCYA4, SCYA5, SCYA22, ANX2, CD83 and UPAR. cDNA array results were confirmed on randomly selected genes using quantitative real-time polymerase chain reaction for three upregulated (CXCR4, IL1B and CD14) and three downregulated (DEK, AF4 and FLI1) genes. Gene expression analysis after differentiation induction may provide a tool to study the roles of DEK, AF4 and FLI1 in cell proliferation and differentiation. To demonstrate the genes that initiate differentiation, sequential gene expression analysis has to be performed during the first 24 h of the differentiation process.
为了确定急性髓系白血病遗传基础进一步研究的潜在靶点，我们研究了与1,25(OH)2D3诱导的HL-60细胞分化相关的基因表达变化。使用包含406个基因（Clontech公司）的阿特拉斯人类血液学滤膜，研究细胞对1,25(OH)2D3（浓度为5×10-8 mol/l）作用24小时和72小时后的基因表达。与未处理的细胞相比，发现43个基因存在表达差异。在两个时间点均下调的基因有：IL2RA、CMYC、NPM、DEK、AF4、FLI1、htlf、MNDA、BCR、IKAROS、BPI和NFAT4。在两个时间点均上调的基因有IL1B、CD14和MCL1。CD55、CD58、IRF2、CREB1、ATF4、RAC1、TIAR、KIAA0053、BAT2、BTK、RCK、EV12B和EDN在24小时时下调，而SPI1、MKK3、BTG1和IL8上调。在72小时时上调的基因有IL1RA、IL2RG、CXCR4、SCYA1、SCYA3、SCYA4、SCYA5、SCYA22、ANX2、CD83和UPAR。使用定量实时聚合酶链反应对三个上调基因（CXCR4、IL1B和CD14）和三个下调基因（DEK、AF4和FLI1）进行随机选择基因的cDNA阵列结果验证。分化诱导后的基因表达分析可能为研究DEK、AF4和FLI1在细胞增殖和分化中的作用提供一种工具。为了证明启动分化的基因，必须在分化过程的最初24小时内进行连续的基因表达分析。
