QUANTITATION AND ARCHIVING OF GENE EXPRESSION

肿瘤恶性细胞群通常不到组织体积的5%。激光捕获显微切割(LCM)提供了一种选择形态相似的恶性细胞的手段，但需要高度灵敏和准确的方法来测量DNA含量、蛋白质和mRNA表达水平，理想情况下都是在相同数量的显微切割细胞中进行的。分子创新公司(MII)将开发一种同时测量LCM细胞基因扩增和表达的精确方法。精确的基因测量是通过设计特定的扩增引物和探针来实现的，它们提供了一种归一化因细胞数量、假基因序列、标本固定、核酸提取、RNA降解和PCR扩增效率而引起的测量波动的方法。提取的核酸不可逆转地与固相材料结合，可以重复分析多个基因(存档)。第一个基因靶点包括雌激素和孕激素受体(ER和PGR)以及c-erbB-2癌基因。人们普遍认为这些基因在乳腺癌的临床治疗中很重要。我们的检测将提供更精确的基因表达测量，定位于与乳腺肿瘤病理相关的特定组织细胞。拟议的商业应用：乳腺癌是女性癌症死亡的主要原因。一种改进的、更精确的检测基因表达水平的方法具有巨大的商业潜力，这些基因表达水平已被证明具有预后和治疗重要性。

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.