Genotoxic effects of diethylstilbestrol on mouse Sertoli TM4 cells using gene expression profiling

Abstract

Diethylstilbestrol (DES), a synthetic estrogen, was examined for genotoxicity in mouse testicular Sertoli TM4 cells using an in vitro micronucleus assay and microarray analysis to clarify the molecular mechanisms underlying the genotoxicity of estrogenic compounds on the male reproductive system. The micronucleus test showed that DES induced genotoxic effects on TM4 cells with S9 activation. Gene expression profiles were studied in DES-treated cells and positive controls, which were cyclophosphamide (CPA)-treated TM4 cells, as compared to the negative controls. In total, 349 and 328 genes were identified as being either up- or down-regulated, with over 2-fold changes, in DES- and CPA-treated TM4 cells, respectively. Biofunction and canonical pathways of differentially expressed genes were analyzed using Ingenuity Pathways Analysis, which were mainly categorized as cellular development and growth/proliferation. In addition, genes related to cell cycle regulation, such as Egr1, Far1, Cd44, Wint16, Sox6, Sor14, Dnmt3a, and Hdac11, were differentially expressed in DES-treated TM4 cells. A gene network analysis was also performed. Comprehensive gene expression profiling of DES-treated TM4 cells provides valuable information to better understand the genotoxic events of estrogenic chemicals in testicular Sertoli cells.