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Updated: Aug 17, 2026
Development of Human Renal Tubular Epithelial Cell Primary Cultures in Monolayers and Three-Dimensional Conditions
Published on: June 13, 2025
Molecular markers of trichloroethylene-induced toxicity in human kidney cells
Lawrence H Lash1, David A Putt, Sarah E Hueni
1Department of Pharmacology, Wayne State University School of Medicine, 540 East Canfield Avenue, Detroit, MI 48201, USA. l.h.lash@wayne.edu
Abstract:
Difficulties in evaluation of trichloroethylene (TRI)-induced toxicity in humans and extrapolation of data from laboratory animals to humans are due to the existence of multiple target organs, multiple metabolic pathways, sex-, species-, and strain-dependent differences in both metabolism and susceptibility to toxicity, and the lack or minimal amount of human data for many target organs. The use of human tissue for mechanistic studies is thus distinctly advantageous. The kidneys are one target organ for TRI and metabolism by the glutathione (GSH) conjugation pathway is responsible for nephrotoxicity. The GSH conjugate is processed further to produce the cysteine conjugate, S-(1,2-dichlorovinyl)-l-cysteine (DCVC), which is the penultimate nephrotoxic species. Confluent, primary cultures of human proximal tubular (hPT) cells were used as the model system. Although cells in log-phase growth, which are undergoing more rapid DNA synthesis, would give lower LD(50) values, confluent cells more closely mimic the in vivo proximal tubule. DCVC caused cellular necrosis only at relatively high doses (>100 muM) and long incubation times (>24 h). In contrast, both apoptosis and enhanced cellular proliferation occurred at relatively low doses (10-100 muM) and early incubation times (2-8 h). These responses were associated with prominent changes in expression of several proteins that regulate apoptosis (Bcl-2, Bax, Apaf-1, Caspase-9 cleavage, PARP cleavage) and cellular growth, differentiation and stress response (p53, Hsp27, NF-kappaB). Effects on p53 and Hsp27 implicate function of protein kinase C, the mitogen activated protein kinase pathway, and the cytoskeleton. The precise pattern of expression of these and other proteins can thus serve as molecular markers for TRI exposure and effect in human kidney.
Insights
Trichloroethylene (TRI) kidney toxicity is complex. Human proximal tubule cells revealed that the metabolite S-(1,2-dichlorovinyl)-L-cysteine (DCVC) induces apoptosis and proliferation at low doses, offering potential biomarkers for TRI exposure.
Area of Science:
- Toxicology
- Human Cell Biology
- Biochemistry
Background:
- Trichloroethylene (TRI) toxicity evaluation is challenging due to multiple target organs, metabolic pathways, and species-dependent variations.
- Human data for TRI toxicity is limited, making human tissue studies advantageous for mechanistic insights.
- The kidneys are a TRI target organ, with nephrotoxicity mediated by the glutathione (GSH) conjugation pathway and its metabolite, S-(1,2-dichlorovinyl)-L-cysteine (DCVC).
Purpose of the Study:
- To investigate the mechanistic effects of DCVC, a TRI metabolite, on human proximal tubular (hPT) cells.
- To identify potential molecular markers for TRI exposure and its effects in human kidney cells.
Main Methods:
- Utilized confluent, primary cultures of human proximal tubular (hPT) cells as a model system.
- Exposed hPT cells to varying doses and incubation times of DCVC.
- Analyzed cellular responses including necrosis, apoptosis, proliferation, and protein expression related to cell death and growth regulation.
Main Results:
- DCVC induced cellular necrosis at high doses (>100 muM) and long incubation times (>24 h).
- Low doses (10-100 muM) and early incubation times (2-8 h) of DCVC triggered apoptosis and enhanced cellular proliferation.
- Significant changes in protein expression involved in apoptosis (Bcl-2, Bax, Apaf-1, Caspase-9, PARP) and cellular growth/stress response (p53, Hsp27, NF-kappaB) were observed.
Conclusions:
- DCVC elicits distinct cellular responses in human kidney cells, including apoptosis and proliferation at low, environmentally relevant doses.
- Observed protein expression patterns serve as sensitive molecular markers for TRI exposure and its specific effects on human kidney cells.
- The findings highlight the utility of human proximal tubular cell cultures for understanding TRI nephrotoxicity mechanisms and identifying biomarkers.
