QUANTITATION AND ARCHIVING OF GENE EXPRESSION

肿瘤恶性细胞群通常占组织体积的5%以下。激光捕获微解剖（LCM）提供了一种选择形态相似的恶性细胞的方法，但需要高度敏感和准确的方法来测量DNA含量，蛋白质和mRNA表达水平，理想情况下，所有这些都在相同数量的微解剖细胞内。Molecular Innovations, incorporated （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定的扩增引物和探针来完成的，这些引物和探针提供了一种使由于细胞数量、假基因序列、标本固定、核酸提取、RNA降解和PCR扩增效率引起的测量波动归一化的手段。提取的核酸与固相物质不可逆结合，可以对多个基因进行系列重复分析（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治疗中被广泛认为是重要的。我们的分析将提供更精确的基因表达测量定位在特定的组织细胞相关的乳腺肿瘤病理。拟议的商业应用：乳腺癌是女性癌症死亡的主要原因。一种改进和更精确的检测基因表达水平的方法已被证明具有预后和治疗的重要性，具有巨大的商业化潜力。

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.