佛波酯诱导K562细胞分化过程中WT1基因表达及其异构体比例变化的研究  

Expression of WT1 gene and its isomer ratio changes during phorbol ester induced differentiation of K562 cell line

作　　者：李晓红[1] 王宏伟[1] 李建兰[1] 朱镭[1] 樊卫平[1] 田彩霞[1] 徐菁[1] 徐永群[1] 

机构地区：[1]山西医科大学第二医院血液病研究所,太原030001

出　　处：《中华血液学杂志》2007年第6期367-370,共4页Chinese Journal of Hematology

摘　　要：目的分析佛波酯(TPA)诱导 K562细胞分化过程中 WT1基因及其四种异构体表达水平及比例变化,探讨 WT1基因不同异构体与细胞分化的关系。方法采用 TPA 诱导 K562细胞分化,以硝基四氮唑蓝(NBT)还原实验及细胞免疫表型 CD9检测判断细胞分化程度;实时荧光定量 RT-PCR技术检测 K562细胞被诱导分化过程中总 WT1基因表达水平,并计算出 WT1(+/+)、WT1(+/-)、WT1(-/+)、WT1(-/-)四种异构体在细胞分化过程中的比例变化。结果 TPA 诱导 K562细胞分化过程中 NBT 阳性率及 CD9表达阳性率均有显著增加(P<0.05)。WT1相对表达水平由分化前(4.67±1.11)×10^(-3),降到(1.67±0.45)×10^(-3)(P<0.05),随后回升,96 h达(4.64±1.53)×10^(-3);四种异构体比例变化不一致,WT1(+/+)比例由0 h的(39.65±19.46)%降至96 h的(15.25±7.27)%,而 WT1(-/-)异构体由0 h的(15.38±11.34)%上升至96 h的(37.60±11.90)%,另外两种异构体比例变化无显著性差异。结论 TPA 诱导 K562细胞分化过程中总 WT1表达水平24 h内迅速下降,随后回升。分化前异构体以 WT1(+/+)为主,而分化后以 WT1(-/-)为主,提示 WT1基因可能通过调节四种异构体的比例而发挥抑制分化或促进分化的作用。Objective To explore the changes in expression of WT1 gene and ration of its isomers during phorbol ester （ TPA ） induced differentiation of leukemia cell line K562 by fluorescence quantitative RT-PCR and analysis the relationship between different isomers and hematogenic cell differentiation. Methods The degree of cellular maturation were verified by NBT reduction test and immunopbenotyping. Expression of WT1 gene was determined by fluorescence quantitative RT-PCR during differentiation of K562 cell line. The relative ratio of the four splicing variants WT1 （ ＋/＋ ） , WT1 （ ＋/- ）, WT1 （ -/＋ ）, WT1 （ -/- ） were calculated. Results During the differentiation of K562 cell, the NBT reduction rate and the CD9 positive rate both increased significantly （ P 〈 0.05 ）. The expression of WT1 gene decreased immediately to （ 1.67 ＋ 0.45 ） × 10 ^-3 from （4.67 ± 1.11 ） × 10^ -3, and then increased again to （4.64± 1.53 ） × 10 ^-3 at 96 hours. The ratio of WT1 （ ＋ / ＋ ） was decreased gradually, from 0 hour （ 39.65 ±19.46 ） % to 96 hour （ 15.25 ±7.27） %. While the ratio of WT1 （ - / - ） was increased, from 0 hour （ 15.38 ± 11.34） % , to 96 hour （ 37.60 ± 11.90） %. The other two isomers ratios did not change significantly. Conclusion During the TPA induced differentiation of K562 cell, there are two high expression levels of WT1 gene. Before differentiation, the majority is WT1 （ ＋ / ＋ ） , and after differentiation, is WT1 （ - / - ）. It indicates that WT1 gene may activate or inhibit cell differentiation by regulating the ratio of its four splicing variants.

关 键 词：K562细胞 细胞分化 基因 WTl 

分 类 号：R733.7[医药卫生—肿瘤]