丁酸钠诱导K562细胞红系分化基因表达谱分析

Effect of Sodium Butyrate on Erythroid Lineage by Expressional Profiles Microarray in K562 Cells

目的分析丁酸钠(NaB)诱导K562细胞后的差异表达基因,筛选出与红系分化相关基因,探索NaB的作用机制。方法应用Affymetrix公司的人类全基因组HG-U133Plus2芯片,检测0.5mmol/LNaB诱导K562细胞48h的珠蛋白基因变化及差异表达基因,筛选出与红系分化相关基因。应用反转录(RT)-PCR检测ε、γ、β、δ4个珠蛋白基因及2个红系分化相关基因KLF1及KLF3的表达,验证基因芯片结果。结果表达谱基因芯片结果分析显示:总探针组54675个芯片中,阳性表达数23175(42.4%),阴性表达数30693(56.1%)。信号比对数值(SLR)结果显示,表达上调(SLR>1)433个,包含340个不同的基因;表达下调(SLR<-1)171个,包含144个不同的基因。生物学功能分类发现所涉及的基因主要包括参与细胞及大分子代谢、细胞信号、生物学调节、细胞发育及细胞增殖等。7个珠蛋白中ε、Aγ、Gγ、β、δ珠蛋白基因表达均上调,而α及ζ珠蛋白基因无明显改变;表达上调或下调且与红系分化相关的有ALAS2、EGR1、LMO2、RUNX1、KIT、GYPA、GATA-2、KLF1及KLF3等29个基因。RT-PCR验证基因芯片结果可靠。结论NaB诱导K562细胞向红系分化;NaB可能通过调节ALAS2、EGR1、LMO2、RUNX1、KIT、GYPA、GATA-2、KLF1及KLF3等与红系分化相关的基因调控γ珠蛋白合成。

Objective Identification of the differently expressed genes on human leukemic K562 cells induced with sodium butyrate （NaB） by expressional profiles microarray, screening an associated novel gene related with erythroid lineage and investigation the pharmacology effects of NaB. Methods HG - U133A Plus 2 （affymetrix） expressional profiles microarray was used to detect the differential expression genes with 0.5 mmol/L NaB - induced K562 cells. Bioinformatics was used to analyze globin gene and to screen an associated novel gene related with erythroid differentiation in detected results. The GeneChip results confirmed by reverse transcriptase - polymerase chain reaction （RT- PCR） detecting. Results The expression of 23 175 （42.4%） probe sets were present and 30 693 （56.1%） absent in 54 675 total probe sets by expressional profiles mieroarray in K562 cells treated with NaB. Six hundred and four differential expression transcriptons detected by signal log ratio （SLR）, were altered by at least 2 - fold. Of these 604 transcripts, 433 up - regulated（ SLR 〉 1 ） which represented 340 genes and 171 down - regulated （ SLR 〈 - 1 ） represented 144 genes. Significantly different expression genes could regulate mostly cellular and macromolecule metabolic proeess, cell communication, regulation of biological process, cellular developmental process and the cell proliferation, using information available from the GeneOntology database. The ε、Aγ ,Gγ ,β and δ globin gene were up- regulated, whereas α and ζ globin gene were not marked change in 7 globin genes. Twenty - nine genes related with erythroid differentiation were up - regulated or down- regulated include ALAS2, EGR1, LMO2, RUNX1, KIT, GYPA, GATA -2, KLF1, KLF3 and so on. RT- PCR detecting showed that thet GeneChip results were reliable. Conclusions NaB has pharmacology effects to differentiation to the erythroid lineage in K562 cells;the differently expressed genes related with erythroid differentiation such as ALAS2, EGR1, LMO2, RUNX1, KIT, GYPA, GATA -2, KLF1, KLF3 would provide a way on the potential pharmacology effects to induce the erythroid differentiation in K562 cells.