正常人骨髓不同细胞群中WT1基因表达及其异构体比例研究  

WT1 Gene Expression and Its Isoform Ratio in Different Cell Subsets of Normal Human Bone Marrow

作　　者：徐菁[1] 王宏伟[1] 李晓红[1] 朱镭[1] 张丽[1] 张帆[1] 覃艳红[1] 杨涛[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：《中国实验血液学杂志》2007年第3期603-606,共4页Journal of Experimental Hematology

基　　金：山西省自然科学基金资助项目(编号20051102);山西省留学归国人员基金资助项目(编号2006035)

摘　　要：本研究探讨WT1基因在正常人骨髓不同分化阶段造血细胞中的表达程度及异构体间比例关系。采用实时荧光定量RT-PCR方法检测18例正常人骨髓标本中CD34+CD38-、CD34+CD38+、CD15+CD11b+、CD33+CD14+、CD20+CD5-和CD20-CD5+细胞群中总WT1、WT1(+17AA)及WT1(+KTS)的表达。结果表明:与K562细胞相比,CD34+CD38-、CD34+CD38+、CD15+CD11b+和CD33+CD14+细胞群中均存在WT1基因的低表达,且依次逐渐降低;在CD20+CD5-和CD20-CD5+细胞群中未检测到WT1基因表达。在分化程度较低的CD34+CD38-和CD34+CD38+细胞群中以+17AA,+KTS及WT1(+/+)异构体为主,而较成熟的CD15+CD11b+和CD33+CD14+细胞群中以-17AA、-KTS及WT1(-/-)异构体为主。结论:在正常人骨髓细胞中WT1基因的表达随着细胞分化程度提高而降低,造血细胞可能通过其4种异构体比例的调整而发挥抑制或促进分化的作用。The Wilms＇tumor gene （WT1） is a transcription factor involved in tumorigenesis, especially in leukemogenesis. However, the role of WT1 expression in nonmalignant hematopoietic cells remains unclear. Furthermore, due to alternative splicing at two sites： 17 amino acid residues of exon 5 （ ＋ 17AA） and 3 amino acid residues （ ＋ KTS） between exons 9 and 10, WT1 gene has four main isoforms （ 17AA ＋/KTS ＋, 17AA ＋/KTS-, 17AA-/KTS ＋, 17AA-/KTS- , abbreviation： ＋/＋, ＋/-,-/＋ ,-/-） . The isoforms probably existed in hematopoietic cells, which make the research more complex. The aim of study was to elucidate the expression and its isoforms of WT1 gene in different cell subsets of healthy bone marrow donors. The fluorescence RT-PCR detection system was established to measure the expressions of full-length WT1, WT1 （ ＋ 17AA） and WT1 （ ＋ KTS） in CD34^＋ CD38^-（ stem cell） , CD34^＋ CD38^＋（ progenitor cell）, CD15^＋CD11b^＋ （ granulocyte）, CD33^＋ CD14^＋ （ monocyte）, CD20^＋ CD5^- （B-lymphocyte） and CD20^- CD5^＋ （ T- lymphocyte ） subsets from 18 normal human bone marrow samples. The results showed that WT1 expressed in CD34^＋CD38^-, CD34^＋ CD38^＋ , CD15^＋CD11b^＋ and CD33^＋ CD14^＋ , but not in CD20^＋ CD5^- and CD20^- CD5^＋ subsets. The highest expression was in CD34^＋ CD38^- , but decreased gradually in CD15^＋ CD11b^＋ and CD33^＋ CD14^＋ subsets. WT1 （ ＋ 17AA） , WT1 （ ＋ KTS） and WT1 （ ＋ / ＋ ） isoforms were predominant in CD34^＋ CD38^- and CD34^＋ CD38^＋ primitive subsets, while in CD15^＋ CD11b^＋ and CD33^＋ CD14^＋ the dominant isoforms were WT1 （ -17AA）, WT1 （-KTS） and WT1 （-/-）. It is concluded that the expression of WT1 in normal bone marrow decreases gradually with cell differentiation. Hematopoietic cells may adjust the ratios of WT1 isoforms to inhibit or promote cell differentiation.

关 键 词：骨髓 WT1基因 WT1基因异构体 基因表达 

分 类 号：R733.7[医药卫生—肿瘤]