EDAG-1在白血病和淋巴瘤细胞株中的表达  

Expression of EDAG-1 Gene in Human Leukemia and Lymphoma Cell Lines

作　　者：周颖[1] 许望翔[2] 詹轶群[2] 李长燕[2] 徐诚望[2] 郑红[1] 李菲菲[1] 杨晓明[2] 汪思应[1] 

机构地区：[1]安徽医科大学基础医学院病理生理学教研室,安徽合肥230032 [2]军事医学科学院放射医学研究所,北京100850

出　　处：《癌症》2004年第11期1238-1243,共6页Chinese Journal of Cancer

基　　金：国家自然科学基金(No.30170464);国家杰出青年科学基金(No.30025018);国家高科技发展计划基金(No.2001AA221321);安徽省生物医药产业化专项基金(No.a01303006);安徽省人才开发基金(No.2002Z035)~~

摘　　要：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: Embryonic development associated gene 1 (EDAG 1), located at chromosome 9q22, is specially expressed in hematopoietic cells, and related to the regulation of hematopoietic system. This study was designed to explore relationship between pathogenesis of leukemia,lymphoma and EDAG 1 through analyzing the structure of EDAG 1 coding region, and its expression in these cell lines. METHODS: Fifteen leukemia and lymphoma cell lines, HEL, K562, HL 60, Namalwa, Raji, J111, Jurkat, HuT 78, MEG 01, U937, 6T CEM, HPB ALL, KG 1a, THP 1, and DAMI, were selected to observe the expression of EDAG 1 by reverse transcriptase polymerase chain reaction (RT PCR), EDAG 1 cDNA coding fragments (1.5 kb) were purified to construct the corresponding recombinant plasmid. Then, the plasmid was sequenced to analyze mutation of the coding region. The expression of EDAG 1 protein, and mRNA in these cell lines were detected by Western blot, and Northern blot; the rearrangement and amplification of EDAG 1 genome in these cell lines were detected by Southern blot. RESULTS: EDAG 1 mRNA and protein were highly expressed in erythroleukemia cell lines (K 562, HEL), megakaryoblast leukemia cell lines (DAMI, MEG 01), and T cell leukemia cell line (Jurkat), while no gene mutation was found in coding region, no amplification and rearrangement of genome was detected in these cell lines. EDAG 1 was absent in HL 60 cell line, and rearranged in HuT 78 cells. CONCLUSION: EDAG 1 may relate with pathogenesis of erythroleukemia and megakaryoblast leukemia; its coding region may have no relation with the mechanism of its activation.

关 键 词：白血病 淋巴瘤 EDAG-1 重排 表达 

分 类 号：R733[医药卫生—肿瘤]