ORGANIZATION AND CONTROL OF GENETIC MATERIAL IN PLASMACYTOMAS

浆细胞瘤中遗传物质的组织和控制

• 批准号: 3774338
• 负责人: J F MUSHINSKI
• 金额: --
• 依托单位: DIVISION OF CANCER BIOLOGY AND DIAGNOSIS
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3774338
• 关键词: 3T3 cells; B lymphocyte; RNA splicing; Retroviridae; athymic mouse; cell cycle proteins; cell differentiation; cell growth regulation; chemical carcinogen; chemical carcinogenesis; chromosome translocation; human tissue; isozymes; lymphoma; molecular cloning; neoplasm /cancer genetics; oncogenes; petroleum oil; phorbols; plasma cell neoplasm; protein kinase C; protooncogene; structural genes; viral carcinogenesis; virus genetics

Our research goal is to understand the genes whose altered structure or expression play critical roles in malignancy, cell growth and normal differentiation. We are concentrating on the study of the expression of a group of known oncogenes: abl, bcl-2 and myc, as well as potential oncogenes: Pvt-1, protein kinases C (PKC) and cyclins, in mouse hematopoietic tissues and tumors. Deregulated expression of myc secondary to chromosomal translocations has been shown to be one essential element in the series of genetic alterations that are involved in oil-induced plasmacytomas in BALB/c mice. Some of these myc-activating translocations occur 200-300 kb 3' of c-myc in a region designated Pvt-l. We have shown that Pvt- l and c-myc are co-amplified in a series of B-cell lymphomas and expressed at a very high level. Pvt-l expression utilizes alternative splicing of numerous exons, including one that may be as far as l Mb 3' of c-myc. We have recently identified three areas of structural similarity between the Pvt-l genes of mouse and man. This evolutionary conservation suggests strongly that Pvt-1 is a gene of considerable importance to normal cellular metabolism. The ABL-MYC retrovirus, that expresses v-abl and c-myc, rapidly induces i.p. plasmacytomas in BALB/c mice. This transformation also works in vitro if the virus is used to infect suspensions of lymphoid cells that are subsequently transplanted into syngeneic mice. This works even if the cells are early B lymphocyte precursors, i.e., pro-B or pre-B cells. If the mice are immunized before induction of plasmacytomas, 50% of them develop tumors that produce antibodies that are directed toward the immunogen. This technology has been used to produce monoclonal antibodies to several protein and peptide antigens. These results suggest that the most efficient B-cell transformation by ABL-MYC occurs in mature, committed B lymphocytes. Eight isozymes of the PKC family have been shown by us to be expressed in a cell-type specific fashion in hematopoietic cells and cell lines. Expression vectors that stably overexpress all of these isoforms have been prepared. Overexpression of certain of these PKCs in NIH3T3 cells has proved to be transforming in vitro and tumorigenic in nude mice. Similarly, overexpression of these PKC isoforms in a mouse myeloid cell line, 32D, has been shown to impart to these cells the ability to differentiate into macrophages when exposed to phorbol esters. Cyclins B1, D1, D2 and D3 have been cloned and used to identify the chromosome that bears these genes. Expression of these cyclins in hematopoietic tumors has been shown to be cell-type specific.

我们的研究目标是了解结构或结构发生改变的基因 表达在恶性肿瘤、细胞生长和正常情况中发挥关键作用 差异化。我们专注于研究a的表达 一组已知的致癌基因：abl、bcl-2 和 myc，以及潜在的 致癌基因：小鼠中的 Pvt-1、蛋白激酶 C (PKC) 和细胞周期蛋白 造血组织和肿瘤。 myc 次级表达失调 已证明染色体易位是一个基本要素 在与石油相关的一系列基因改变中 BALB/c 小鼠浆细胞瘤。其中一些 myc 激活易位 出现在c-myc的200-300kb 3'处指定为Pvt-1的区域中。我们已经展示了 Pvt-l 和 c-myc 在一系列 B 细胞淋巴瘤中共同扩增，并且 表达了很高的水平。 Pvt-l 表达利用替代 大量外显子的剪接，其中一个可能长达 l Mb 3' c-myc。我们最近发现了三个结构相似的领域 小鼠和人的 Pvt-l 基因之间的关系。这种进化保守 强烈表明 Pvt-1 是一个对于 正常的细胞代谢。 表达 v-abl 和 c-myc 的 ABL-MYC 逆转录病毒可快速诱导 ip BALB/c 小鼠浆细胞瘤。这种转变在体外也有效 如果病毒被用来感染淋巴细胞悬浮液 随后移植到同系小鼠中。即使 细胞是早期 B 淋巴细胞前体，即 pro-B 或前 B 细胞。如果 小鼠在诱导浆细胞瘤前进行免疫，其中50% 产生的肿瘤会产生针对 免疫原。该技术已用于生产单克隆抗体 多种蛋白质和肽抗原。这些结果表明 ABL-MYC 最有效的 B 细胞转化发生在成熟、 定型B淋巴细胞。 我们已证明 PKC 家族的八种同工酶在 造血细胞和细胞系中的细胞类型特异性方式。 稳定过表达所有这些亚型的表达载体已被开发出来 准备好了。 NIH3T3 细胞中某些 PKC 的过度表达 被证明在体外具有转化性并且在裸鼠中具有致瘤性。 同样，这些 PKC 同工型在小鼠骨髓细胞中的过度表达 细胞系 32D 已被证明可以赋予这些细胞以下能力： 当暴露于佛波酯时分化为巨噬细胞。 细胞周期蛋白 B1、D1、D2 和 D3 已被克隆并用于鉴定 携带这些基因的染色体。这些细胞周期蛋白的表达 造血肿瘤已被证明具有细胞类型特异性。