【作者】 朱永进；

【导师】 薛永权；

【作者基本信息】 苏州大学 ， 内科血液学， 2006， 博士

【摘要】 目的和意义白血病是严重危害人类健康的造血系统恶性疾病,常伴有非随机的细胞遗传学异常,其中最常见的是染色体易位。在急性白血病中约65%的患者伴有特征性的染色体易位,染色体易位的受累基因大多为编码转录因子或受体的酪氨酸激酶基因,编码的融合蛋白干扰细胞增殖、凋亡和分化的正常调控途径,在恶性血液病发病机制中起着重要作用。因此,检测恶性血液病非随机的染色体异常,分离染色体异常受累基因,并进一步研究这些基因的功能及其与恶性血液病发生、发展的关系,对理解恶性血液病的发病机制,寻找有效的诊断标志和治疗方法都具有重要意义。近年来,与染色体11p15异常相关的恶性血液病病例不断增多,至今已发现17种与恶性血液病相关的染色体11p15异常,其中15种染色体易位的靶基因均为位于11p15的NUP98基因。本课题旨在通过对一例伴t(3;11)(q29q13;p15)的急性杂合性白血病的研究,定位克隆t(3;11)(q29q13;p15)染色体易位的受累基因,并对NUP98的伙伴基因及形成的融合基因进行功能初探,以深化我们对NUP98相关融合基因致白血病的发生、发展的认识,从而有助于指导白血病的诊断、预后评估和治疗。方法我们对一例伴t(3;11)(q29q13;p15)的原发性急性杂合性白血病进行了深入研究,充分利用当前人类基因组研究所取得的大量信息,采取了定位候选基因克隆策略(positional candidate gene approaches),定位克隆t(3;11)(q29q13;p15)染色体易位的受累基因,进而对NUP98相关融合蛋白的致白血病机制进行了初步的研究。第一部分:首先通过常规的分子-细胞遗传学方法检测出t(3;11)( q13;p15)染色体易位,并将11号染色体上的断裂点初步定位于11p15的NUP98基因3’端;接着通过cDNA末端快速扩增技术(Rapid Amplification of cDNA Ends, RACE)发现了位于3q29的NUP98伙伴基因NRG(因当时GeneBank无此基因的详细信息,我们姑且命名为NUP98 Related Gene);为了解释RACE结果与核型检测不符之处,我们选取了位于3q21和位于3q29的两个BAC克隆来分析3号染色体复杂异常情更多还原

【Abstract】 ObjectiveLeukemia is a kind of malignant tumor of hematopoietic system doing severely harm to human health, always with non-random cytogenetic abnormalities, among which the most frequency is chromosomal translocation. It can be seen in 65% acute leukemia patients. Mostly targed genes of chromosomal translocation are transcriptional factor or trosine-protein kinase, resulting in dysregulative pathwalys of cell proliferation, apoptosis, differentiation and playing an important role in leukemogenesis. Therefore, detection of the recurrent chromosomal translocations, molecular cloning and functional study of the involved genes will shed light on deciphering the molecular mechanisms of leukemogenesis, provide diagnostic markers and allow the development of new therapeutic strategies. In recent years, chromosome aberration involving 11p15 has been reported increasingly. To date at least 17 kinds have been identified. Almost all 11p15 translocations targed the NUP98 gene located at chromosome 11p15. Our work is to provide considerable insight into the mechanisms of NUP98-related fusion genes in leukemia through molecular cloning and functional study of the fusion genes by the chromosome translocation t(3;11)(q29q13;p15) in a hybrid acute leukemia patient Furthermore,it will help to diagnose, guide risk-directed therapy for leukemia.MethodsWe investigated an adult patient with the chromosome translocation t(3;11)(q29q13;p15) in de novo hybrid acute leukemia. With positional candidate gene approaches, we characterized the genes involved in that translocation and subsequently studied the function of the fusion protein in leukemogenic mechanism.Part I: Firstly, through molecular genetic-cytogenetics, we identified a new chromosomal translocation t(3;11)( q13;p15) and located chromosome breakpoint at the NUP98 gene at 11p15. Using Rapid Amplification of cDNA Ends (RACE), we更多还原