Expression cloning of anticancer drug-sensitivity-regulatory molecules in gynecologic malignancy

本研究利用我们首次从妇科肿瘤细胞系中建立的多种抗癌耐药亚克隆，对抗癌耐药相关基因进行了分子克隆。此外，我们还对我们建立的耐药亚系的抗癌药物敏感性的调控机制进行了分析。首先，我们对从不同耐药亚克隆中提取的RNA进行了差异显示。实验还没有完成。采用差异显示的方法。筛选出乙醛脱氢酶、T细胞白血病相关基因、细胞色素C基因和白细胞介素13受体基因等与抗癌药物敏感性/耐药性相关的候选基因。我们现在正在对候选基因进行功能分析，虽然实际上我们通过对已建立的耐药亚系的分析发现了许多耐药机制的发现，但我们没有足够的时间在国际期刊上发表这些结果。我们对100多个耐药亚克隆进行了对其他抗癌药物的交叉耐药性、已知耐药相关基因的表达(定量Taq-Man RT-PCR)、免疫细胞化学和流式细胞术的分析。我们正在利用DNA微阵列技术研究1,176个基因在耐药癌细胞中的差异表达。我们正在与已经开发出抗癌药物的制药公司一起寻找候选药物，以恢复耐药细胞的药物敏感性。

In this study, molecular cloning of anticancer drug-resistance-related genes were performed by using various anticancer drug-resistant subclones we have originally established from gynecological cancer cell lines. Moreover, analyses of regulatory mechanisms of anticancer drug-sensitivity in the drug-resistant sublines we established were also examined.First, we have been applied differential display procedure to RNA extracted from the various drug-resistant subclones. The experiments have not been finished yet. By the differential display method. several candidate genes related to anticancer drug-sensitivity/resistance such as aldehyde dehydrogenase, T-cell leukemia related genes, cytochome C gene and interleukin-13 receptor gene were selected. We are now under investigation of functional analyses of the candidate genes.Although in fact we have found lots of discovery in drug-resistance mechanisms by analyses of the established drug-resistant sublines, we did not have enough time to publish these results on international journals. On more than one hundred of resistant subclone, we analyzed them by cross-resistance to other anticancer drugs, expression of known drug-resistance-related genes (quantitative Taq-Man RT-PCR), immunocytochemistry and flow cytometry. Chromosomal analyses were also examined.We are investigating differential expressions of 1,176 genes in the drug-resistance cancer cells by DNA microarray technology. Together with pharmaceutical company which have developed anticancer drugs, we are searching candidate drugs to recover drug-sensitivity in the drug-resistant cells.