Selection of drucrs against ovarian cancer and trial of discovering new molecular targets using gene expression profiles

为了阐明在卵巢癌的致癌步骤和获得耐药性中起关键作用的基因，我们使用寡核苷酸微阵列系统研究了大约6，000个基因的表达。采用聚类分析技术分析基因表达模式，标准化正常卵巢组织中的基因表达水平。在书面知情同意的情况下，将来自手术标本的卵巢癌组织冷冻在-80 ℃，并从组织中提取mRNA。卵巢癌组织与正常卵巢组织的基因表达谱明显不同。然而.各组织学亚型之间无显著差异。97个基因在癌组织中普遍表达上调，227个基因表达下调。在这些基因中，细胞角蛋白18、Am-1=Evil、HER 3、keratin 19、ear-2、βtubulin、GSTπ、c-erb-B2、nm 23、BRCA 2、p57、IGFBP5.6、Brush-1和TGFB 1BP被认为是分子靶点 ...更多信息 开发新的抗癌药物。紫杉醇是抗卵巢癌的高效药物之一，其作为有丝分裂纺锤体毒物，即，紫杉醇促进微管蛋白的组装并稳定它们，防止解聚。β-tsubulin在卵巢癌组织中表达上调的结果为紫杉醇治疗卵巢癌提供了理论依据。然后，我们研究了人卵巢癌细胞KF 28的基因表达谱，制备了KF 2B的耐药亚克隆KFr 13（对顺铂耐药）和KF 28 TX和KFr 13 TX（对紫杉醇耐药）。顺铂抗性克隆显示与谷胱甘肽解毒途径、糖酵解/糖原生成、转酮醇酶和多胺合成酶相关的基因高表达。紫杉醇耐药克隆高表达MDR和semaphorinE等多个耐药基因。比较临床因素与本研究中鉴定的基因表达水平，将有助于开发新的分子靶向药物，并阐明耐药获得的机制。少

To clarify genes that play critical roles for carcinogenesis steps and acquisition of drug resistance of ovarian cancer, we studies approximately 6,000 gene expressions using an oligonucleotide microarray system. Gene expression patterns were analyzed by clustering technique, which standardized the gene expression level in normal ovarian tissues. With written informed consent, ovarian cancer tissues from surgical specimens were frozen at-80 C and mRNA was extracted from the tissues. Clustered gene expression profiles of ovarian cancer tissues were clearly different from those of normal tissues. However. there were no significant differences among each histological subtype of cancers. The expression levels of 97 genes were commonly up-regulated in cancer tissues and those of 227 genes were down-regulated. Among these genes, cytokeratin18, Am-1=Evil, HER3, keratin19, ear-2, βtubulin, GSTπ, c-erb-B2, nm23, BRCA2, p57, IGFBP5.6, Brush-1 and TGFB1BP were thought to become a molecular target … More of developng new anti-cancer drugs. One of highly effective drugs against ovarian cancer, paclitaxel, acts as a mitotic spindle poison, i.e., paclitaxel promotes assembly of tubulins and stabilizes them, preventing depolymerization. Our results that βtsubulin was up-regulated in all cancer specimens gave a theoretical evidence to the application of paclitaxel against ovarian cancer treatment. Then, we studied gene expression profiles of human ovarian cancer cell, KF28, Drug-resistant subclones of KF2B were prepared, KFr13, which is resistant to cis-platinum and, KF28TX and KFr13TX, which are resistant to paclitaxel. The cis-platinum-resistant clone showed the high expression of genes related with depoisoning pathway through glutathione, with glycolysis/ glycogenesis, with transketolase and with polyamine synthesis enzymes. The paclitaxel-resistant clones highly expressed multiple drug resistant genes, MDR and semaphorinE, etc. Comparison of clinical factors with expression levels of genes identified in this study will help to develop new molecular target drugs and to clarify mechanism of the acquisition of drug resistance. Less