QUANTITATION AND ARCHIVING OF GENE EXPRESSION

基因表达的定量和存档

• 批准号: 2869468
• 负责人: JOHN Charles GERDES
• 金额: $ 12.88万
• 依托单位: XTRANA, INC.
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-05-17 至 2001-05-16
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2869468
• 关键词: breast neoplasms; cytogenetics; estrogen receptors; gene expression; human genetic material tag; neoplasm /cancer genetics; neoplastic cell; nucleic acid quantitation /detection; polymerase chain reaction; progesterone receptors; protooncogene; technology /technique development

Tumor malignant cell populations frequently occupy less than 5% of the tissue volume. Laser capture microdissection (LCM) provides a means of selecting malignant cells of similar morphology, but highly sensitive and accurate methods are needed for measuring DNA content, protein and mRNA expression levels ideally all within the same small number of microdissected cells. Molecular Innovations, incorporated (MII) will develop a precise method for simultaneous measurement of gene amplification and expression from LCM cells. Precise gene measurement is accomplished through the design of specific amplification primers and probes that provide a means of normalizing measurement fluctuations due to cell number, pseudogene sequences, specimen fixation, nucleic acid extraction, RNA degradation, and PCR amplification efficiency. Extracted Nucleic Acids are bound irreversibly to a solid phase material and can be repeatedly analyzed in series for multiple genes (archiving). The first gene targets include Estrogen and Progesterone Receptors (ER and PgR) and the c-erbB-2 oncogene. These genes are well accepted as important in the clinical management of breast cancer. Our assay will provide for more precise gene expression measurement localized in the specific tissue cells relevant to the breast tumor pathology. PROPOSED COMMERCIAL APPLICATIONS: Breast cancer is a leading cause of cancer deaths in women. An improved and more precise method of detecting gene expression levels that have been shown to be of prognostic and therapeutic importance has great commercialization potential.

肿瘤恶性细胞群通常占组织体积的5%以下。激光捕获显微切割（LCM）提供了一种选择类似形态的恶性细胞的方法，但需要高度灵敏和准确的方法来测量DNA含量，蛋白质和mRNA的表达水平，理想的是所有在相同的少量显微切割细胞。分子创新公司（MII）将开发一种精确的方法，用于同时测量LCM细胞的基因扩增和表达。精确的基因测量是通过设计特异性扩增引物和探针来实现的，这些引物和探针提供了一种使测量波动标准化的方法，该测量波动是由于细胞数量、假基因序列、标本固定、核酸提取、RNA降解和PCR扩增效率引起的。提取的核酸与固相材料不可逆地结合，并且可以重复地连续分析多个基因（存档）。第一个基因靶点包括雌激素和孕激素受体（ER和PgR）和c-erbB-2癌基因。这些基因在乳腺癌的临床管理中被广泛接受为重要的。我们的测定将提供定位于与乳腺肿瘤病理学相关的特定组织细胞中的更精确的基因表达测量。建议的商业应用：乳腺癌是女性癌症死亡的主要原因。一种改进的和更精确的检测基因表达水平的方法，已被证明是预后和治疗的重要性，具有很大的商业化潜力。