Drug metabolism enzyme expression and activity in primary cultures of human proximal tubular cells

Toxicology. 2008 Feb 3;244(1):56-65. doi: 10.1016/j.tox.2007.10.022. Epub 2007 Nov 4.


We previously catalogued expression and activity of organic anion and cation, amino acid, and peptide transporters in primary cultures of human proximal tubular (hPT) cells to establish them as a cellular model to study drug transport in the human kidney [Lash, L.H., Putt, D.A., Cai, H., 2006. Membrane transport function in primary cultures of human proximal tubular cells. Toxicology 228, 200-218]. Here, we extend our analysis to drug metabolism enzymes. Expression of 11 cytochrome P450 (CYP) enzymes was determined with specific antibodies. CYP1B1, CYP3A4, and CYP4A11 were the only CYP enzymes readily detected in total cell extracts. These same CYP enzymes, as well as CYP3A5 and possibly CYP2D6, were detected in microsomes from confluent hPT cells, although expression levels varied among kidney samples. In agreement with Western blot data, only activity of CYP3A4/5 was detected among the enzyme activities measured. Expression of all three glutathione S-transferases (GSTs) known to be found in hPT cells, GSTA, GSTP, and GSTT, was readily detected. Variable expression of three sulfotransferases (SULTs), SULT1A3, SULT1E, and SULT2A1, and three UDP-glucuronosyltransferases (UGTs), UGT1A1, UGT1A6, and UGT2B7, was also detected. When examined over the course of cell growth to confluence, expression of all enzymes was generally maintained at readily measurable levels, although they were often lower than in fresh tissue. These results indicate that primary cultures of hPT cells possess significant capacity to metabolize many classes of drugs, and can be used as an effective model to study drug metabolism.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Blotting, Western
  • Cell Growth Processes / drug effects
  • Cells, Cultured
  • Chromatography, Liquid / methods
  • Coumarins / chemistry
  • Coumarins / metabolism
  • Coumarins / pharmacology
  • Cytochrome P-450 Enzyme System / metabolism
  • Enzymes / metabolism*
  • Felodipine / administration & dosage
  • Felodipine / metabolism
  • Female
  • Glucuronosyltransferase / metabolism
  • Glutathione Transferase / metabolism
  • Humans
  • Isoenzymes / metabolism
  • Kidney Tubules, Proximal / cytology
  • Kidney Tubules, Proximal / enzymology*
  • Kidney Tubules, Proximal / metabolism
  • Male
  • Mass Spectrometry / methods
  • Metabolic Detoxication, Phase I
  • Metabolic Detoxication, Phase II
  • Microsomes / drug effects
  • Microsomes / enzymology
  • Microsomes / metabolism
  • Middle Aged
  • Pharmaceutical Preparations / administration & dosage
  • Pharmaceutical Preparations / metabolism*
  • Sulfotransferases / classification
  • Sulfotransferases / metabolism
  • Time Factors


  • Coumarins
  • Enzymes
  • Isoenzymes
  • Pharmaceutical Preparations
  • Cytochrome P-450 Enzyme System
  • Glucuronosyltransferase
  • Glutathione Transferase
  • Sulfotransferases
  • Felodipine