Quantitative Analysis of UGT2B28 mRNA Expression by Real-Time RT-PCR and Application to Human Tissue Distribution Study

Shuji      Ohno; Shizuo      Nakajin

Abstract

We recently established a method for quantitative determination of human catalytically active UDPglucuronosyltransferases (UGTs) other than UGT2A1 and UGT2B28 by real-time reverse transcription-polymerase chain reaction (RT-PCR), and applied the method to an exhaustive analysis of localization in various human tissues. We report here an additional quantitative determination method targeting UGT2B28. To date, there have been no reports on the distribution of UGT2B28 mRNA expression in human tissues based on quantitative determination. Human UGT2B28 was clearly detected in the breast and adipose tissue. UGT2B28 expression in the breast was comparatively low, about 1.6% of GAPDH mRNA levels, and was less than 5% of normalized (against GAPDH) UGT2B7 and 2B10 mRNA expression levels in the liver. Although the UGT2B28 has 97% identity with UGT2B11 at the cDNA sequence level, the primers constructed for UGT2B28 did not detect UGT2B11. In addition, significant expression of UGT2B11 was detected in the liver, breast and kidney, and was clearly different from the distribution of UGT2B28. Therefore, we conclude that the real-time RT-PCR method established here is very specific for human UGT2B28 isozyme.

Keywords: Human UGTs, mRNA expression, quantitative analysis, real-time RT-PCR, tissue distribution, UGT2B28, adrenal gland, skeletal muscle, primers, bilirubin