Effects of zinc ions and HrasG12V-activated signaling on gene expression of rat embryonic fibroblasts maintained in serum-free media

To identify intracellular events affected by HrasG12V and zinc ion availability, gene expression profiling was performed with Rat-1·RasValA1, a stably transfected cell line where oncogenic mutant HrasG12V is expressed under the control of an IPTG-inducible promoter. Rat-1·RasValA1 cells were treated for 24 h with or without 5 mM IPTG in serum-free DMEM in the presence or absence of ZnCl2 supplementation at 3 microM. Total RNA was purified for gene expression profiling with microarray. In the study presented here assessing the effects of HrasG12V and zinc ions on gene expression profiles, Rat-1·RasValA1 cells, which express HrasG12V in an IPTG-inducible manner, were used. They were maintained in serum-free DMEM and exposed to the following 4 different conditions for 24 h: IPTG(-) ZnCl2(-), IPTG(+) ZnCl2(-), IPTG(-) ZnCl2(+), and IPTG(+) ZnCl2(+). Expression profiles of a total of ~2,000 genes were acquired.