Objective: Understanding gene expression variations by using RNA transcript analysis methods during hepatic viral protein interactions with the IFN pathway in hepatic cell lines has recently gained importance. One of the most powerful techniques in gene expression quantification is quantitative real-time RT-PCR. Reference genes used as normalizer in this method may be affected across various experimental conditions or treatments. Hence, in the present study, the influence of IFN-a treatment on the mRNA levels of common reference genes including ACTB, GAPDH, TBP, HPRT1 and HMBS was evaluated in Huh-7 or HepG2 cell lines.
Methods: Cells were treated with different concentrations of IFN-a. Then, using geNorm and NormFinder programs, we evaluated the expression stabilities of the above prominent reference genes in three sample groups that included each hepatic cell line and the total data sets.
Results: HPRT-1 and GAPDH were the most stable reference genes in the Huh-7 cell line, whereas ATCB, HMBS and GAPDH were the most stable in the HepG2 cell line. TBP was one of the least stable reference genes in the three studied groups.
Conclusion: This investigation will provide appropriate reference genes for standardization of quantitative real-time PCR data in an IFN-a stimulated model of hepatocyte cell lines.