Establishment and comprehensive gene analysis of cells derived from 3-dimensional radial flow bioreactor culture system

为建立用于药物疗效监测和评价的体外人工器官模型系统，以生化检测为主要手段，对一种新颖的径向流型生物反应器(RFB)三维培养系统的有效性进行了检验。在此条件下，对药物代谢酶、耐药相关分子、细胞凋亡相关分子等基因表达的调控进行了综合分析，并建立了相关数据库。经三维培养，肝癌细胞(FLC系列)表达的CYP分子种类超过预期。另一方面，药物转运和代谢的分子物种基本没有变化。此外，肝癌细胞还增加了血浆蛋白的产生。通过三维培养，肿瘤细胞控制形成三维结构所必需的细胞骨架相关基因和参与细胞间连接的基因群的表达。许多获得的新观点将使知识产权发生。当比较单层培养和RFB培养的FLC细胞的药物反应时，RFB培养的细胞显著减弱了抗癌药物的作用。在RFB培养中对鳞状细胞癌的治疗也证实了药效的下降。现在，它正在分析表达基因的变化。由于肝细胞癌和鳞状细胞癌的短期治疗，药物主要影响应激诱导基因的表达，而不影响耐药基因或代谢基因的表达。由于以往报道的领域较少，因此对这些结果很有兴趣并进行了分析，在此基础上，我们想对RFB主导的基因表达进行综合分析，并建立数据库。

To establish of in vitro artificial organ model system for monitoring and evaluating the efficacy of medicine, usefulness of an original three dimensional culture system of radial flow type bioreactor (RFB) was examined using mainly biochemical determination in the drug responses. Under these conditions, the control of gene expressions such as drug metabolizing enzymes, drug resistance related- and apoptosis related-molecules was analyzed comprehensively from the result and a database of them was built. The liver cancer cells (FLC series) expressed CYP molecular species above the expectations by the three dimensional cultivation. On the other hand, the molecular species about the drug transportation and metabolism didn't change roughly. Moreover, the liver cancer cells increased the production of plasma proteins. By the three dimensional cultivation, the tumor cells controlled the expression of the cytoskeleton related-genes which are necessary for forming the three dimensional structure and the gene group which participates in the intercellular junction. A lot of gotten new-observations will make an intellectual property occur. When comparing the cellular drug-response of the FLC cells between a monolayer culture and a RFB cultivation, the cells cultured in RFB weakened the efficacy of the anticancer drug remarkably. The decline of the medicine effect was also confirmed in the treatment of the squamous cell carcinoma in the RFB cultivation. Now, it is analyzing the changes of the expressed genes. Because of the short term treatment of the liver cell cancer and the squamous cell carcinoma, the medicines affected mainly on the expression of stress-induced genes rather than that of genes of either drug-resistance or metabolism. Because the report is one with scarce field in the past, these results have an interest and are analyzed.Based on this result, we want to comprehensively analyze the gene expression which the RFB led and to make a database.