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

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，重要的是确定表征乳腺肿瘤发生的起始步骤的分子改变。应用SAGE（SerialAnalysisofGene Expression）技术比较了正常乳腺上皮细胞和DCIS（DuctalCarcinoma In-Situ，a precursor of invasive BC）乳腺上皮细胞的基因表达谱。该技术鉴定了HIN-1（High In Normal-1），一种在正常乳腺上皮细胞中高度表达的新型分泌蛋白，但在>90%的原发性浸润前和浸润性BC和BC细胞系中丢失。HIN- 1在这些早期肿瘤中失活的高频率表明它作为一种新的肿瘤抑制因子。HIN-1也在正常肺、胰腺、前列腺和唾液腺中高度表达，所有器官都含有分支上皮。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，并确定其在早期BC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。将使用腺病毒和质粒载体或通过暴露于纯化的重组体将HIN-1引入BC细胞系中。HIN-1和使用集落生长测定和细胞周期进程的流式细胞术分析的效果进行分析。具体目标2将在三维培养系统中探索HIN-1表达对乳腺上皮细胞分支形态发生的影响。具体目标3将使用用四环素诱导型HIN-1载体转染的BC细胞系解决HIN-1抑制裸鼠中肿瘤细胞生长的能力。因此，这些研究的结果将确定HIN-1作为潜在肿瘤抑制基因的作用，并且由于它在早期BC中经常失活，因此可能为预防性干预提供极好的靶点。