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背景与目的:EDAG-1(embryonicdevelopassociatedgene1,EDAG-1)定位于9q22,在造血细胞特异表达,并与造血调控关系密切。本研究通过检测EDAG-1在白血病细胞和淋巴瘤细胞中的表达和编码区基因结构,探讨EDAG-1与白血病和淋巴瘤发病的关系。方法:选用白血病或淋巴瘤细胞株共15种,采用RT-PCR法检测表达EDAG-1基因的细胞株,并回收该基因cDNA编码区片段,构建相应的重组质粒并测序分析编码区突变情况。通过Northernblot和Westernblot分别确证EDAG-1mRNA和蛋白在白血病和淋巴瘤细胞株中的表达情况,利用Southernblot检测EDAG-1在白血病细胞和淋巴瘤细胞株中基因重排和扩增情况。结果:红系(K-562、HEL)、巨核系(DAMI、MEG-01)、Jurkat细胞在mRNA、蛋白水平均高表达EDAG-1,但其编码区结构未发现异常,也没有EDAG-1基因组的扩增和重排。发现HL-60细胞缺失EDAG-1,HuT78细胞EDAG-1发生重排。结论:EDAG-1可能与红系、巨核系白血病的发病机制有关,该基因激活的机制可能与编码区突变无关。
BACKGROUND & OBJECTIVE: EDAG-1 (EDAG-1) is located on 9q22 and is specifically expressed in hematopoietic cells and is closely related to the regulation of hematopoiesis. In this study, the expression of EDAG-1 in leukemia cells and lymphoma cells and the gene structure of coding region were detected to explore the relationship between EDAG-1 and leukemia and lymphoma. Methods: A total of 15 leukemia or lymphoma cell lines were selected. The cell lines expressing EDAG-1 gene were detected by RT-PCR. The cDNA coding region of the gene was recovered and the corresponding recombinant plasmids were constructed. The expression of EDAG-1 mRNA and protein in leukemia and lymphoma cell lines was confirmed by Northern blot and Western blot respectively. The gene rearrangement and amplification of EDAG-1 in leukemia cells and lymphoma cell lines were detected by Southern blot. Results: EDAG-1 was highly expressed at mRNA and protein levels in erythroid (K-562, HEL), megakaryocyte (DAMI, MEG-01) and Jurkat cells, but no abnormalities in the coding region and no EDAG-1 Genome amplification and rearrangement. It was found that EDAG-1 was deleted in HL-60 cells and EDAG-1 in HuT78 cells was rearranged. Conclusion: EDAG-1 may be involved in the pathogenesis of erythroid and megakaryocytic leukemias. The mechanism of this gene activation may not be related to mutation in the coding region.