CD44-P185HER2 INTERACTION IN OVARIAN CANCER PROGRESSION

卵巢癌是妇科恶性肿瘤死亡的主要原因， 美国的 尤其是卵巢癌， 女性生殖道最致命的肿瘤，并继续成为 是女性癌症患者死亡的主要原因。 肿瘤转移 当卵巢增大导致患者 症状（晚期III-IV）。 生物因素影响 卵巢肿瘤细胞迁移和植入远处的能力 腹膜腔内的位置仍然不清楚。 拟议 研究中，我们计划检验CD 44亚型-p185 HER 2 与细胞骨架的相互作用在促进 肿瘤细胞粘附，运动， 在人类卵巢癌进展过程中的增殖和生长。 这 这一假设显然对传播有重要影响， 人类卵巢癌的进展。具体来说，我们计划使用一个 生物化学、免疫学和分子生物学 阐明结构和功能关联的技术 在CD 44 s-p185 HER 2复合物和细胞骨架之间。 我们还将 使用体外诱变构建CD 44 s缺失（或位点- 定向的）突变体，其缺少某些部分（或替换某些氨基）， 酸）的细胞质结构域的结合所需的 细胞骨架 这些突变多肽将在卵巢癌细胞中表达， 肿瘤细胞及其结构变化将与CD 44相关， p185 HER 2介导的致癌特性（例如细胞粘附，运动， 细胞增殖和生长）。在建议的第二部分，我们 计划鉴定与人类免疫缺陷病毒相关的CD 44变体（CD 44 v）亚型， 使用RT-PCR、DNA序列分析和原位杂交检测卵巢癌 杂交方法我们将独特的变异外显子克隆到标准CD 44中， cDNA，并在CD 44 v阴性（p185 HER 2- 阳性）卵巢上皮细胞。 然后这些CD 44 v转染子将 分析其功能特性（例如，CD 44 v-p185 HER 2 相互作用、粘附、肿瘤细胞运动、增殖和生长）。 我们还计划下调CD 44 v-p185 HER 2介导的致癌信号 通过使用Shc和/或Grb 2的显性负突变体。 这些突变 多肽将在卵巢肿瘤细胞中表达， 结构变化将与各种致癌特性相关。 最后，针对“融合”蛋白的特异性抗体含有 将制备CD 44变体外显子（与卵巢癌细胞相关）。 CD 44 v亚型与p185 HER 2在卵巢癌中的共表达 将在卵巢癌进展期间分析癌。 我们 我认为，从这项建议中获得的新信息可能 建立CD 44 v-p185 HER 2复合物作为重要的肿瘤标志物， 早期检测和评估致癌潜力，以及允许 开发新的药物靶点来抑制肿瘤细胞运动， 增殖/生长。

Ovarian cancer is the leading cause of gynecologic malignancy deaths in the United States. In particular, ovarian carcinomas appears to be the most lethal tumor of the female genital tract and continues to be the major cause of mortality in female cancer patients. Metastases from the ovary are common by the time the ovarian enlargement leads to patient symptoms (at advanced stages III-IV). The biological factors affecting the ability of ovarian tumor cells to migrate and implant at distant locations within the peritoneal cavity remain unclear. In the proposed research, we plan to test the hypothesis that the CD44 isoform-p185HER2 interaction with the cytoskeleton plays an important role in promoting oncogenic signals required for tumor cell adhesion, motility, proliferation and growth during human ovarian cancer progression. This hypothesis clearly has important implications for the spread and progression of human ovarian cancers. Specifically, we plan to use a variety of biochemical, immunological and molecular biological techniques to elucidate the structural and functional association between the CD44s-p185HER2 complex and the cytoskeleton. We will also employ in vitro mutagenesis to construct CD44s deletion (or site- directed) mutants that lack certain portions (or replace certain amino acids) of the cytoplasmic domain required for the binding of the cytoskeleton. These mutants polypeptides will be expressed in ovarian tumor cells and their structural changes will be correlated with CD44s- p185HER2-mediated oncogenic properties (e.g. cell adhesion, motility, cell proliferation and growth). In the second part of the proposal, we plan to identify CD44 variant (CD44v) isoforms associated with human ovarian cancers using RT-PCR, DNA sequence analyses and in situ hybridization. We will clone the unique variant exons into standard CD44 cDNA and express these constructs in CD44v-negative (p185HER2- positive) ovarian epithelial cells. These CD44v transfectants will then be analyzed for their functional properties (e.g. CD44v-p185HER2 interaction, adhesion, tumor cell motility, proliferation and growth). We also plan to downregulate CD44v-p185HER2-mediated oncogenic signals by using dominant-negative mutants of Shc and/or Grb2. These mutant polypeptides will be expressed in ovarian tumor cells and their structural changes will be correlated with various oncogenic properties. Finally, specific antibodies against the "fusion" proteins containing CD44 variant exons (related to ovarian cancer cells) will be prepared. The co-expression of certain CD44v isoforms with p185HER2 in ovarian carcinomas will be analyzed during ovarian cancer progression. We believe that the new information obtained from this proposal may establish the CD44v-p185HER2 complex as an important tumor marker for early detection and evaluation of oncogenic potential, as well as allow the development of new drug targets to inhibit tumor cell motility and proliferation/growth.