Evaluation of variation of glucuronidation activity and substrate specificity caused by difference in the proportion of expression level of UGT isozimes in living cells

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The aim of this study was to clarify the factor of gap between UGT activity evaluated by in vitro and in vivo assay. Then we have developed an assay method to determine UGT activity in living cells (as near in vivo model). The method is as follows: LLC-PK1 cells were incubated with substrate for 2 hours on ice. After incubating, the cells were incubated on water bath at 37 degree Celsius for enzymes and transporters working. And then metabolites were extracellularly excreted and quantified.Furthermore, we established stable HeLa cell line expressing UGT1A9 and introduced UGT2B4 into it. UGT1A9 activity of HeLa cells expressing UGT2B4 reduced. This result indicated the protein-protein interaction between the UGTs.