Changes in the disposition of estradiol 17-D-glucuronide (E 2 17G), a substrate of the organic anion-transporting polypeptide family (Oatp) and multidrug resistance-associated protein 2 (Mrp2), were examined in livers of male Wag/Rij rats that were injected with CC531 cells intraportally to induce metastatic tumors (n ϭ 5) or with phosphate-buffered saline for shamoperated controls (n ϭ 4). Multiple indicator dilution, singlepass liver perfusions revealed extremely high influx clearances of [ 3 H]E 2 17G (Ͼ190 ml/min) in both groups. In recirculating liver perfusions, [ 3 H]E 2 17G decayed monoexponentially in the reservoir perfusate, and the total (9.19 Ϯ 1.33 versus 8.18 Ϯ 0.94 ml/min) and biliary (4.94 Ϯ 1.07 versus 4.60 Ϯ 0.86 ml/min) clearances were similar in both groups (P Ͼ 0.05). The metabolic clearance of E 2 17G was higher in the tumor group (4.60 Ϯ 0.64 versus 3.23 Ϯ 0.23 ml/min, P Ͻ 0.05). E 2 3S17G, the 3-sulfate metabolite, whose identity was confirmed by mass spectrometry, appeared only in bile and not perfusate. Liver microsomal incubations of E 2 3 35 S17G and [ 3 H]estrone sulfate revealed similar sulfatase activities between the tumor and sham livers, albeit the activities were much lower for E 2 3 35 S17G. Oatp1a1 and Oatp1b2 protein expression in liver membrane fragments was reduced by 42% and 38%, respectively, whereas that of cytosolic estrogen sulfotransferase (Sult1e1) was significantly increased (41%) with tumor (P Ͻ 0.05). All of the observations were captured by modeling. From modeling, we showed that reduction of the high influx clearance (546 to 283 ml/min) failed to lower the total clearance of E 2 17G, whereas up-regulation of Sult1e1 increased the E 2 17G sulfation clearance (2.56 to 3.69 ml/min) in livers with metastatic tumors.The liver mediates both the metabolism and excretion of drugs. Molecules gain access into hepatocytes where the machinery for vectorial transport and biotransformation exists. At the basolateral/sinusoidal membrane, unbound molecules in plasma are taken up by hepatocytes via organic anion/ cation transporters and/or passive diffusion. In the cell, the molecules can be metabolized by various phase I and phase II enzymes. Finally, drug precursors and metabolites are either secreted into bile by the ATP binding cassette transporters, the multidrug resistance-associated protein (MRP) 2, P-glycoprotein, and breast cancer resistance protein (BCRP), or effluxed back into the blood stream by MRP3 and MRP4.Liver tumor metastasis is a progressive invasion of malig-