Cell Biology and Toxicology

, Volume 31, Issue 4–5, pp 221–230 | Cite as

Evaluation of CYP3A4 inhibition and hepatotoxicity using DMSO-treated human hepatoma HuH-7 cells

  • Yitong LiuEmail author
  • Thomas J. Flynn
  • Menghang Xia
  • Paddy L. Wiesenfeld
  • Martine S. Ferguson
Original Article


A human hepatoma cell line (HuH-7) was evaluated as a metabolically competent cell model to investigate cytochrome P450 3A4 (CYP3A4) inhibition, induction, and hepatotoxicity. First, CYP3A4 gene expression and activity were determined in HuH-7 cells under three culture conditions: 1-week culture, 3-week culture, or 1 % dimethyl sulfoxide (DMSO) treatment. HuH-7 cells treated with DMSO for 2 weeks after confluence expressed the highest CYP3A4 gene expression and activity compared to the other two culture conditions. Furthermore, CYP3A4 activity in DMSO-treated HuH-7 cells was compared to that in a human hepatoma cell line (HepG2/C3A) and human bipotent progenitor cell line (HepaRG), which yielded the following ranking: HepaRG > DMSO-treated HuH-7 >> HepG2/C3A cells. The effects of three known CYP3A4 inhibitors were evaluated using DMSO-treated HuH-7 cells. CYP3A4 enzyme inhibition in HuH-7 cells was further compared to human recombinant CYP3A4, indicating similar potency for reversible inhibitors (IC 50 within 2.5-fold), but different potency for the irreversible inhibitor. Next, induction of CYP3A4 activity was compared between DMSO-treated HuH-7 and HepaRG cells using two known inducers. DMSO-treated HuH-7 cells yielded minimal CYP3A4 induction compared to that in the HepaRG cells after 48-h treatments. Finally, the cytotoxicity of five known hepatotoxicants was evaluated in DMSO-treated HuH-7, HepG2/C3A, and HepaRG cells, and significant differences in cytotoxic sensitivity were observed. Overall, DMSO-treated HuH-7 cells are a valuable model for medium- or high-throughput screening of chemicals for CYP3A4 inhibition and hepatotoxicity.


HuH-7 cells DMSO CYP3A4 Inhibition Induction Hepatotoxicity 



The authors thank Dr. Michael F. Santillo for image processing and critical review of the paper. The authors thank Ms. Shelia PughBishop for technical assistance. This research was supported by the US Food and Drug Administration. This research report is not an official US Food and Drug Administration guidance or policy statement. No official support or endorsement by the US Food and Drug Administration is intended, nor should it be inferred.

Conflict of interest

The authors declare no conflicts of interest.


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Copyright information

© US Government 2015

Authors and Affiliations

  1. 1.Division of Toxicology, Office of Applied Research and Safety Assessment, Center for Food Safety and Applied NutritionUS Food and Drug AdministrationLaurelUSA
  2. 2.Division of Pre-Clinical Innovation, National Center for Advancing Translational SciencesNational Institutes of HealthBethesdaUSA
  3. 3.Division of Public Health Informatics & Analytics, Office of Analytics and Outreach, Center for Food Safety and Applied NutritionUS Food and Drug AdministrationCollege ParkUSA

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