Real-time quantitative PCR analysis of gene expression in human colon cancer cells treated with LPS

Relative mRNA levels of 55 genes were evaluated with the SYBR Green qPCR assays after LPS treatment for 8 h. Overall design: Human colon cancer cells (COLO 225) were purchased from ATCC. The cells in triplicate were treated with various concentrations of LPS for 8 h. RNA isolation and cDNA synthesis were performed as described previously. Relative mRNA levels were evaluated with the SYBR Green qPCR assays using Bio-Rad reagents. BCL2 mRNA was selected as the internal reference for qPCR analysis for this cell type. 1% DMSO treatment was the sample control. The 2-ΔCT and 2-ΔΔCT method of relative quantification was used to determine the fold change in expression.