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: 3752050
• 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/3752050
• 关键词: 3T3 cells; B lymphocyte; athymic mouse; cell cycle proteins; cell differentiation; cell growth regulation; chromosome translocation; gene mutation; genetic regulation; genetic strain; immunocytochemistry; lymphoma; molecular cloning; monoclonal antibody; neoplasm /cancer genetics; oncogenes; phorbols; plasma cell neoplasm; protein kinase C; protooncogene; tumor suppressor genes

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其结构或结构发生变化的基因 基因表达在肿瘤、细胞生长和正常生长中起着关键作用 差异化。我们正在集中精力研究A的表达方式 一组已知的癌基因：abl、bcl2和myc，以及潜在的 癌基因：PVT-1、蛋白激酶C(PKC)和细胞周期蛋白 造血组织和肿瘤。Myc继发性表达的解除调控 染色体易位已被证明是一个基本因素 在石油引起的一系列基因改变中 BALB/c小鼠的浆细胞瘤。其中一些激活myc的易位 C-myc的200-300kb的3‘端出现在一个被命名为pvt-L的区域。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 以非常高的水平表达。PVT-L表达利用替代性 多个外显子的拼接，包括一个可能远至L Mb 3‘的外显子 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基因之间的差异。这种进化守恒 强烈提示Pvt-1是一个对 正常的细胞代谢。 表达v-abl和c-myc的ABL-myc逆转录病毒迅速诱导 IP地址。BALB/c小鼠的浆细胞瘤。这种转化在体外也有效。 如果病毒被用来感染淋巴细胞的悬液， 随后移植到同基因小鼠体内。这是有效的，即使 细胞是早期的B淋巴细胞前体，即前B细胞或前B细胞。如果 这些小鼠在浆细胞瘤诱导前进行免疫，其中50%的小鼠 发展产生抗体的肿瘤，抗体针对的是 免疫基因。这项技术已被用于生产单抗。 对几种蛋白质和多肽抗原。这些结果表明， ABL-MYC最有效的B细胞转化发生在成熟的， 承诺的B淋巴细胞。 我们已经证明了PKC家族的八种同工酶在 造血细胞和细胞系中的一种特定的细胞类型。 稳定过度表达所有这些异构体的表达载体已经被 准备好了。其中某些PKC在NIH3T3细胞中过表达 被证明在体外转化并在裸鼠体内致瘤。 同样，这些PKC亚型在小鼠髓系细胞中过表达 品系32D已被证明赋予这些细胞 当接触佛波酯时，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 带有这些基因的染色体。这些细胞周期蛋白在人乳腺癌细胞中的表达 造血肿瘤已被证明是细胞类型特异性的。