House Keeping Genes and Expression Analysis using a Taqman Human Endogenous Control Plate in Human Hepatic Tissue

Aim: Quantification of mRNA transcripts using real time RT PCR may be compounded by variation within one tissue type, leading to misinterpretation of the expression profiles of target genes. To identify a suitable house keeping gene for use in hepatic gene expression analysis of patients with haemochromatosis and hepatitis C.

Method: A TaqMan™ Human Endogenous Control Plate evaluating the expression of eleven select housekeeping genes was used to validate internal controls for the analysis of gene expression in human liver tissue.

Results: Eight house keeping genes, including beta actin and GAPDH were up regulated in

haemochromatosis and hepatitis C. Beta actin displayed a 6.3–8.92 fold higher expression, and GAPDH, a 1.02–3.35 fold higher expression. Transferrin receptor (TfR) had the greatest ÄCt range, from -2.21 to +5.36. The control with the least variation in ÄCt levels was cyclophilin with values of -2.72, -2.76, -2.26 from patients with hepatitis C and two haemochromatosis patients respectively.

Conclusion: This observation demonstrates that there is wide variation in the levels of transcription among house keeping genes in liver tissue of patients with hepatitis C and haemochromatosis.

Cyclophilin was expressed at constant levels in all samples.