Molecular analysis of B-cell malignant lymphoma-related oncogenes and its clinical application for diagnosis and treatment.

B细胞恶性淋巴瘤相关癌基因的分子分析及其临床诊断和治疗应用。

• 批准号: 13671091
• 负责人: HOSOKAWA Yoshitaka
• 金额: $ 2.62万
• 依托单位: Aichi Cancer Center
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671091/
• 关键词: Malignant lymphoma; Chromosome translocation; apoptosis; BCL6

1) BCL6. The BCL-6/LAZ3 gene encodes a zinc-finger transcriptional repressor and is located at the breakpoint of the 3q27-associated translocations that occur most frequently in non-Hodgkin's lymphomas (NHLs To examine cell responses and target genes related to the BCL-6 signaling pathway, we established Ba/F3 pro-B cells carrying a human BCL-6 transgene that is inducible under control of the lactose operon. Using a CDNA array hybridization technique, we found that the induced BCL-6 protein can downregulate the expressions of the genes, cyclin A2, chemokine receptor CXCR4, and insulin-like growth factor binding protein-4 (IGFBP-4) in the Ba/F3 cells. Northern blot analysis established that the expressions of these genes were indeed downregulated by the induced BCL-6 protein but in a somewhat different manner. The induced BCL-6 protein also inhibited cell proliferation of Ba/F3 cells. These findings strongly suggest that three key genes, namely cyclin A2, CXCR4 and IGFBP-4 may play a ro … More le in the downstream of the BCL-6 signaling pathway during B - lymphoid differentiation.2) API2-MALT1. t(11;18)(q21;q21) is a characteristic chromosomal translocation in mucosa-associated lymphoid tissue (MALT) type lymphoma, and this translocation results in chimeric transcript of API2 (Apoptosis Inhibitor 2, also known as c-IAP2) - MALT1 (Mucosa-Associated Lymphoma Translocation gene 1). To identify proteins that bind API2-MALT1 chimeric protein, we employed coimmunoprecipitation and SDS PAGE analysis, followed by liquid chromatography-electrospray ionization tandem mass spectrometry. As a result, Smac, Htra2, and TRAF2 were identified as API2-MALT1-binding proteins. Immunoprecipitation analysis demonstrated that ectopically expressed API2-MALT1 protein indeed binds to these endogeneous proteins. Furthermore, API2-MALT1 protein significantly inhibited Smac-promoted apoptosis in UV irradiated HeLa cells. These data strongly suggest that API2-MALT1 can block apoptosis, at least in part, by inhibiting Smac-mediated apoptotic pathway. Less

1)BCL6。BCL-6/LAZ 3基因编码锌指转录抑制因子，位于非霍奇金淋巴瘤（NHL）中最常发生的3q 27相关易位的断点处。为了检查与BCL-6信号传导途径相关的细胞反应和靶基因，我们建立了携带人BCL-6转基因的Ba/F3 pro-B细胞，该转基因在乳糖操纵子的控制下是可诱导的。利用cDNA阵列杂交技术，我们发现诱导的BCL-6蛋白能下调Ba/F3细胞中细胞周期蛋白A2、趋化因子受体CXCR 4和胰岛素样生长因子结合蛋白4（IGFBP-4）等基因的表达。北方印迹分析证实，这些基因的表达确实被诱导的BCL-6蛋白下调，但方式略有不同。诱导表达的BCL-6蛋白对Ba/F3细胞的增殖也有抑制作用。这些发现有力地表明，三个关键基因，即cyclin A2，CXCR 4和IGFBP-4，可能发挥作用。 ...更多信息 le在B -淋巴分化过程中BCL-6信号通路的下游。2）API 2-MALT 1。t（11; 18）（q21; q21）是粘膜相关淋巴组织（MALT）型淋巴瘤中的特征性染色体易位，并且该易位导致API 2（凋亡抑制剂2，也称为c-IAP 2）-MALT 1（粘膜相关淋巴瘤易位基因1）的嵌合转录物。为了鉴定结合API 2-MALT 1嵌合蛋白的蛋白，我们采用免疫共沉淀和SDS PAGE分析，然后通过液相色谱-电喷雾串联质谱。结果，Smac、Htra 2和TRAF 2被鉴定为API 2-MALT 1结合蛋白。免疫沉淀分析表明，异位表达的API 2-MALT 1蛋白确实结合这些内源性蛋白。此外，API 2-MALT 1蛋白显着抑制Smac促进的细胞凋亡在紫外线照射的HeLa细胞。这些数据有力地表明，API 2-MALT 1可以阻断凋亡，至少部分，通过抑制Smac介导的凋亡途径。少

项目成果

細川好孝: "イカロスによるリンパ球分化制御と造血器腫瘍"Molecular Medicine. 38. 784-789 (2001)

Yoshitaka Hosokawa：“伊卡洛斯的淋巴细胞分化控制和造血肿瘤”分子医学 38. 784-789 (2001)。

細川好孝: "DNAチップとその応用"血液の事典 朝倉書店. (印刷中). (2003)

细川芳孝：“DNA 芯片及其应用”血液朝仓书店百科全书（2003 年出版）。

Hosokawa et al.: "Target genes downregulated by the BCL6/LAZ3 oncoprotein in mouse Ba/F3 cells"Biochem.Biophy.Res.Commun.. 3. 563-568 (2001)

Hosokawa 等人：“小鼠 Ba/F3 细胞中 BCL6/LAZ3 癌蛋白下调的靶基因”Biochem.Biophy.Res.Commun.. 3. 563-568 (2001)

細川好孝: "Ikaros gene family"生化学「ことば」. 74. 257 (2002)

细川芳孝：《Ikaros基因家族》《生物化学》74。257（2002）。

Yonezumi M, et al.: "Detection of API2-MALT1 chimeric gene in extranodal and nodal marginal zone B cell lymphoma by RT-PCR and LA-PCR analyses"Br.J.Haematol.. 115. 588-594 (2001)

Yonezumi M 等：“通过 RT-PCR 和 LA-PCR 分析检测结外和结外边缘区 B 细胞淋巴瘤中的 API2-MALT1 嵌合基因”Br.J.Haematol.. 115. 588-594 (2001)