CHARACTERIZATION OF ANGIOTENSIN BINDING IN CELLS TRANSFECTED WITH MAS ONCOGENE

一种可能的血管紧张素受体(来自人类的癌基因)是 联合感染NIH-3T3细胞及致瘤性检测 从人表皮样癌中提取DNA。这些报告表明 这种基因在转染后的重排导致了 人类Mas基因的激活和随后的转化(焦点 形成能力)。注射这些 转化成裸鼠的细胞形成肿瘤。关于转型的分析 3T3细胞提示蛋白质的合成增加 能够结合血管紧张素Ⅱ和血管紧张素Ⅲ。免疫荧光分析 抗肽-针对第一和第二细胞外产生的抗体 Mas多肽的结构域支持存在 移植的3T3细胞的受体合成。这些加在一起， 结果提示，mass基因产物参与了细胞的生长和 扩散。Mas蛋白是否具有血管紧张素功能 受体或作为刺激结合的生长因子的结合部位 血管紧张素，还是血管紧张素与细胞的结合增加 由MA存在引起的生长是细胞类型选择性的，而不是 完全理解。因此，这项建议的目标是：(一) 血管紧张素基因在NIH-3T3细胞中的结合特性 人类Mas基因，(Ii)确定Mas基因的调控作用 细胞的生长和增殖，(Iii)确定细胞是否对 大量表达是特定于细胞类型或机制的，以及(Iv) Mas基因表达与血管紧张素受体活性的相关性 妊娠和未妊娠兔的子宫。学生(包括毕业生和 本科生)将会参与到这项工作的各个方面 求婚。

A putative receptor for angiotensin (mas oncogene from human) was detected by cotransfection and tumorigenicity assay of NIH-3T3 cells with DNA isolated from Human epidermoid carcinoma. These reports suggested that rearrangement of this gene after transfection resulted in the activation of the Human mas gene and subsequent transformation (Foci forming ability) of the transfected cells. Injection of these transformed cells into nude mice form tumors. Analysis of transformed 3T3 cells suggest that there is an increased synthesis of a protein capable of binding Ang II and Ang III. Immunofluorescence analysis using antipeptide-antibodies generated against the 1st and 2nd extracellular domains of the Mas polypeptide support the presence of an increase in receptor synthesis in transplanted 3T3 cells. Taken together, these results suggest that the mas gene product participates in cell growth and proliferation. Whether the Mas protein functions as an angiotensin receptor or as a binding site for growth factors stimulating the binding of angiotensin, or whether the increased binding of angiotensin and cell growth resulting from the presence of Mas is cell-type selective is not fully understood. Therefore, the objectives of this proposal are (i) to characterize the binding of angiotensin in NIH-3T3 cells transfected with the human mas gene, (ii) to determine the role of mas gene in regulating cell growth and proliferation, (III) to determine if the cells response to mas expression is cell-type or mechanism specific, and (iv) to correlate mas gene expression and angiotensin receptor activities in uteri of pregnant and non-pregnant rabbits. Students (both graduate and undergraduate) are expected to participate in all aspects of this proposal.