Construction and validation of human cDNA microarray for estimation of endocrine disrupting chemicals (KISTCHIP-400 ver. 1.0)

Abstract

Transcript profiling is a particularly valuable tool in the field of steroid receptor biology, as these receptors are ligand-activated transcription factors and therefore exert their initial effects through altering gene expression in responsive cells. Also, an awareness of endocrine disrupting chemicals (EDCs) and their potential screening methods to identify endocrine activity have been increased. Here we developed an in-house cDNA microarray, named KISTCHIP-400 ver. 1.0, with 416 clones, based on public database and research papers. These clones contained estrogen, androgen, thyroid hormone & receptors, sex hormone signal transduction & regulation, c-fos, c-myc, ps2 gene, metabolism related genes etc. Also, to validate the KISTCHIP-400 ver. 1.0, we investigated gene expression profiles with reference hormones, 10(-8) M 17 beta-estradiol, 10(-7) M testosterone and 10(-7) M progesterone in MCF -7 cell line. As the results, gene expression profiles of three reference hormones were distinguished from each other with significant and identified 33 17 beta-estradiol responsive genes. This study is in first step of validation for KISTCHIP-400 ver. 1.0, as following step transcriptional profile analysis on not only low concentrations of EDCs but suspected EDCs using KISTCHIP-400 ver. 1.0 is processing. Our results indicate that the developed microarray may be a useful, laboratory tool for screening EDCs and elucidating endocrine disrupting mechanism.