Analysis of clock oscillation in cancer tissue for effective chronotherapy to hypoxic area.

为了监测肿瘤的昼夜节律，建立了稳定引入的DBP-luc结肠癌细胞系。我们发现热休克（43C 1h）不影响DBP-luc细胞的节律期。此外，将缺氧反应元件(HRE)-GFP引入DBP-luc细胞，同时监测缺氧和昼夜节律。为了使DBP-luc / HRE-GFP通过单次转染稳定导入细胞系，我们设计并构建了新的基因构建体。建立了模拟肿瘤组织缺氧的三维体外DBP-luc细胞培养体系。

To monitor cancer circadian rhythm, DBPpromoter-luciferase (DBP-luc) stably introduced colon cancer cell line was established. We found heat shock (43C 1h) did not affects rhythm phase of DBP-luc cell. Additionally, hypoxia responsive element (HRE)-GFP was introduced to DBP-luc cell to monitor hypoxia and circadian rhythm simultaneously. To make DBP-luc / HRE-GFP stably introduced cell line by single transfection, we designed and made new gene constructs. We established the 3D in vitro DBP-luc cell culture system which mimics hypoxia of cancer tissue.