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_医学论文.docVIP

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_医学论文 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_医学论文 作者：徐菁, 王宏伟, 李晓红, 朱镭,张丽,张帆,覃艳红,杨涛 【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。 【关键词】 骨髓　WT1基因　 WT1基因异构体　 基因表达 　　WT1 Gene Expression and Its Isoform Ratio in Different Cell Subsets of Normal Human Bone Marrow Abstract 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.