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方面起关键作用的基因，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。用聚类技术分析基因表达谱，对正常卵巢组织的基因表达水平进行标准化。在获得书面知情同意的情况下，将手术标本中的卵巢癌组织在-80℃下冷冻，并从组织中提取mRNA。卵巢癌组织的聚集性基因表达谱明显不同于正常组织。然而。各组织学亚型间差异无统计学意义。癌组织中共有97个基因表达上调，227个基因表达下调。在这些基因中，细胞角蛋白18、Am-1=EVIL、HER3、角蛋白19、EAR-2、β微管蛋白、谷胱甘肽转移酶π、c-erb-B2、nm23、brca2、p57、IGFBP5.6、Brush-1和TGFB1BP被认为是分子靶…更多的是开发新的抗癌药物。紫杉醇是一种高效的抗卵巢癌药物，它是一种有丝分裂的纺锤体毒药，即紫杉醇促进微管蛋白的聚集并稳定它们，防止解聚。我们的研究结果表明，β转氨酶在所有肿瘤标本中均表达上调，为紫杉醇在卵巢癌治疗中的应用提供了理论依据。然后，我们研究了人卵巢癌细胞KF28的基因表达谱，制备了对顺铂耐药的KF2B亚克隆KF13和对紫杉醇耐药的KF28TX和KFr13TX。顺铂抗性克隆中与谷胱甘肽解毒途径、糖酵解/糖合成、转酮醇酶和多胺合成酶相关的基因表达水平较高。紫杉醇耐药克隆高度表达多药耐药基因、多药耐药基因和信号素E等多药耐药基因。将临床因素与本研究确定的基因表达水平进行比较，将有助于开发新的分子靶向药物，阐明耐药的发生机制。较少

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