Deletion mapping of chromosome band 14q32 for isolation and characterization of the putative tumor related gene in human neuroblastoma

本研究利用12个微卫星标记进行洛丢失（LOH）研究，在染色体14 q32区构建了一个缺失图谱，并对14 q32区的12个微卫星标记进行了分析，结果表明，14 q32区的12个微卫星标记均为缺失，其中14 q32区的12个微卫星标记为缺失，而14 q32区的12个微卫星标记为缺失。我们对54例NB患者的肿瘤及相应组织进行了14 q32微卫星标记的PCR-LOH研究。54例中17例（3.1%）在染色体14 q32区存在杂合性缺失，并发现一个约1cM的共同缺失区，位于D14 S62和D14 S987两侧。本研究利用本基金购买的细菌人工染色体（bacterial artificial chromosome，BAG）和/或从我们构建的人类基因组粘粒文库中分离的粘粒克隆进行FISH分析，证实了上述结果，并构建了一个由BAG和粘粒克隆组成的约8 cM的序列准备好的重叠群，该重叠群两侧是D14 S62和D14 S267，用于分离和分析推定的抑癌基因。我们的研究结果支持NB的发生与14 q32上的抑癌基因有关，但NB的发生机制尚需进一步研究。

We are investigating a tumor suppressor gene(s) responsible for neuroblastoma(NB) and we are focusing especially on chromosome arm 14q.As the first step in isolation and characterization of the putative tumor suppressor gene, we constructed a deletion map on chromosome band 14q32 by LOH study using 12 microsatellite markers.In this study, we analyzed 54 tumors and corresponding constitutional tissues of patients with NB.We performed PCR-LOH study using microsatellite markers on 14q32. Seventeen (3 1%) of 54 cases showed loss of heterozygosity on chromosome band 14q32, and an approximately 1-cM region of common deletion was found that was flanked by D14S62 and D14S987. These results were confirmed by FISH analyses using bacterial artificial chromosome (BAG) purchased by this grant and/or cosmid clones isolated from human genomic cosmid library we constructed.Now, we have constructed an approximately 8-cM sequence-ready contig consisting of BAG and cosmid clones that was flanked by D14S62 and D14S267 to isolate and analyze the putative tumor suppressor gene. Our findings support the *istence of putative tumor suppressor gene on 14q32 for the tumorigenesis of NB.Further investigations are required to elucidate the tumorigenesis of NB.