Characterization of HIN1 Putative Tumor Suppressor

HIN1 假定肿瘤抑制因子的表征

• 批准号: 6709218
• 负责人: IAN E KROP
• 金额: $ 1.57万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-03-25 至 2003-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6709218
• 关键词: Adenoviridae; athymic mouse; breast neoplasms; cell cycle; cell proliferation; flow cytometry; mammary gland; neoplasm /cancer genetics; plasmids; protein structure function; recombinant proteins; transfection /expression vector; tumor suppressor genes; tumor suppressor proteins; Adenoviridae; athymic mouse; breast neoplasms; cell cycle; cell proliferation; flow cytometry; mammary gland; neoplasm /cancer genetics; plasmids; protein structure function; recombinant proteins; transfection /expression vector; tumor suppressor genes; tumor suppressor proteins

To develop preventive interventions for breast cancer (BC), it is important to determine the molecular alterations that characterize the initiating steps of breast tumorigenesis. SAGE (Serial Analysis of Gene Expression) was used to compare the gene expression profiles of normal and DCIS (Ductal Carcinoma In-Situ, a precursor of invasive BC) mammary epithelial cells. This technique identified HIN-1 (High In Normal-1), a novel secreted protein that is highly expressed in normal breast epithelial cells, but lost in >90% of primary pre-invasive and invasive BCs and BC cell lines. The high frequency at which HIN- 1 is inactivated in these early tumors suggests that it acts as a novel tumor suppressor. HIN-1 is also highly expressed in normal lung, pancreas, prostate, and salivary gland, all organs containing branching epithelia. This suggests that HIN-1 may be involved in morphogenesis or differentiation. . The overall objective of the proposed research is to characterize the function of HIN-1 and to determine its role in the initiating steps of early BC. Specific aim 1 is to determine the effect of HIN-1 on cell proliferation. HIN-1 will be introduced into BC cell lines using adenoviral and plasmid vectors or by exposure to purified recombinant. HIN-1 and the effects analyzed using colony growth assays and flow cytometric analysis of cell cycle progression. Specific aim 2 will explore the effects of HIN-1 expression on branching morphogenesis of mammary epithelial cells in a three-dimensional culture system. Specific aim 3 will address the ability of HIN-1 to suppress tumor cell growth in nude mice using BC cell lines transfected with a tetracycline inducible HIN-1 vector. The results of these studies will thus determine the role of HIN-1 as a potential tumor suppressor gene and, since it is inactivated frequently in early BC, may provide an excellent target for preventive intervention.

为了开发乳腺癌（BC）的预防性干预措施，确定表征乳腺肿瘤发生步骤的分子改变很重要。使用SAGE（基因表达的序列分析）来比较正常和DCI（导管癌对立）的基因表达谱，这是侵入性BC的前体）乳腺上皮细胞。该技术确定了HIN-1（正常-1中的高），这是一种新型的分泌蛋白质，在正常乳腺上皮细胞中高度表达，但损失了> 90％的原发性前侵入性和侵入性BCS和BC细胞系。在这些早期肿瘤中，Hin-1灭活的高频表明它是一种新型的肿瘤抑制剂。 HIN-1在正常肺，胰腺，前列腺和唾液腺中也高度表达，所有器官含有分支上皮。这表明HIN-1可能参与形态发生或分化。 。拟议研究的总体目标是表征HIN-1的功能，并确定其在BC早期的启动步骤中的作用。具体目的1是确定HIN-1对细胞增殖的影响。 Hin-1将使用腺病毒和质粒向量或暴露于纯化的重组剂中引入BC细胞系。 HIN-1和使用菌落生长测定法和细胞周期进程的流式细胞术分析进行了分析的效果。具体目标2将探索HIN-1表达对三维培养系统中乳腺上皮细胞的分支形态发生的影响。特定的目标3将解决HIN-1使用四环素诱导型HIN-1载体转染的BC细胞系抑制裸鼠肿瘤细胞生长的能力。因此，这些研究的结果将确定HIN-1作为潜在的肿瘤抑制基因的作用，并且由于它在BC早期经常失活，因此可能为预防性干预提供了绝佳的目标。