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改变或 表达在恶性，细胞生长和正常状态中起关键作用 分化。我们专注于研究A的表达 一组已知的癌基因：ABL，BCL-2和MYC以及潜力 癌基因：PVT-1，蛋白激酶C（PKC）和细胞周期蛋白，在小鼠中 造血组织和肿瘤。 MYC次级的放松调节表达 染色体易位已被证明是一个必不可少的元素 在一系列遗传改变中 BALB/C小鼠中的浆细胞瘤。其中一些是MYC激活的易位 在指定PVT-L的区域中发生200-300 kb 3'c-Myc。我们已经显示了 该PVT-L和C-MYC在一系列B细胞淋巴瘤中共扩散 在很高的水平上表达。 PVT-L表达使用替代 许多外显子的剪接，包括可能到达L MB 3'的外显子。 c-myc。我们最近确定了结构相似性的三个领域 在小鼠和人的PVT-L基因之间。这种进化保护 强烈暗示PVT-1是一个重要性的基因 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 I.P. BALB/C小鼠中的浆细胞瘤。这种转变也可以在体外起作用 如果该病毒用于感染淋巴样细胞的悬浮液 随后将其移植到合元小鼠中。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之前先进行免疫，其中50％ 开发产生针对抗体的肿瘤 免疫原。该技术已用于生产单克隆抗体 到几种蛋白质和肽抗原。这些结果表明 ABL-MYC最有效的B细胞转换发生在成熟中， 承诺的B淋巴细胞。 我们已经证明了PKC家族的八个同工酶在 造血细胞和细胞系中的细胞型特异性方式。 稳定过表达所有这些同工型的表达向量已经 准备。 NIH3T3细胞中某些PKC的过表达已 事实证明，裸鼠的体外和肿瘤症正在转化。 同样，这些PKC同工型在小鼠髓样细胞中的过表达 已显示32D线32D可以赋予这些单元的能力 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已被克隆并用于识别 带有这些基因的染色体。这些细胞周期蛋白在 造血肿瘤已显示为细胞型特异性。