蛍光性低酸素誘導因子の相関分析

Abstract

HIF-1α, hypoxia induced factor-1αis regarded as a target for drug development in several diseases such as cancer. For high throughput screening of HIF-1α-targeted drug, we need to determine the activity sensitively and quantitatively. In the present study, we proposed a method of fluorescence correlation analysis on HIF-1α activation in which Co2+ treatment against stable transformants of GFP-labelled HIF-1α mimicked hypoxia. In fluorescence correlation spectroscopy, we observed fluorescence intensity fluctuation within a volume element which fluorescence molecules enter and exit because of the Brownian motion in solution or cytosol. When one-component model was used for the analysis, it was difficult to discriminate diffusion coefficient of active form of HIF-1α with inactive one. In two-component model, however, a fraction of slow moving component, GFP-labelled HIF-1α increased significantly when the transformants were exposed to Co2+. In the case of high throughput screening for HIF-1α-targeted drug with fluorescence correlation spectroscopy, we should use the fraction of the slower moving component to judge the activation.