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

三维径向流生物反应器培养体系的建立及细胞基因综合分析

基本信息

批准号：
17300160
负责人：
OHKAWA Kiyoshi
金额：
$ 8.58万
依托单位：
Jikei University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17300160/
关键词：
Bioreactor Biomaterial Artificial organ Microarray Proteome ロテオーム

项目摘要

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.

为建立监测和评价药物疗效的体外人工器官模型系统，采用以生化测定为主的方法，对原始径向流型生物反应器（RFB）三维培养系统的有效性进行了考察。在此条件下，综合分析药物代谢酶、耐药相关分子和凋亡相关分子等基因表达的调控情况，并建立数据库。通过三维培养，肝癌细胞（FLC系列）表达CYP分子种高于预期。另一方面，药物转运和代谢的分子种类变化不大。此外，肝癌细胞增加了血浆蛋白的产生。通过三维培养，肿瘤细胞控制了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。大量的新观察将会产生知识产权。比较单层培养和RFB培养的FLC细胞的细胞药物反应，RFB培养的细胞明显削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治疗时间较短，药物主要影响应激诱导基因的表达，而不是影响耐药或代谢基因的表达。由于该报告是过去稀缺领域的报告，因此这些结果具有兴趣并进行了分析。在此基础上，我们希望对RFB所导致的基因表达进行综合分析，并建立数据库。