Changes in gene expression in human renal proximal tubule cells exposed to low concentrations of S-(1,2-dichlorovinyl)-l-cysteine, a metabolite of trichloroethylene.
Epidemiology studies suggest that there may be a weak association between high level exposure to trichloroethylene (TCE) and renal tubule cell carcinoma. Laboratory animal studies have shown an increased incidence of renal tubule carcinoma in male rats but not mice. TCE can undergo metabolism via glutathione (GSH) conjugation to form metabolites that are known to be nephrotoxic. The GSH conjugate, S-(1,2-dichlorovinyl)glutathione (DCVG), is processed further to the cysteine conjugate, S-(1,2-dichlorovinyl)-l-cysteine (DCVC), which is the penultimate nephrotoxic species. We have cultured human renal tubule cells (HRPTC) in serum-free medium under a variety of different culture conditions and observed growth, respiratory control and glucose transport over a 20 day period in medium containing low glucose. Cell death was time- and concentration-dependent, with the EC(50) for DCVG being about 3 microM and for DCVC about 7.5 microM over 10 days. Exposure of HRPTC to sub-cytotoxic doses of DCVC (0.1 microM and 1 microM for 10 days) led to a small number of changes in gene expression, as determined by transcript profiling with Affymetrix human genome chips. Using the criterion of a mean 2-fold change over control for the four samples examined, 3 genes at 0.1 microM DCVC increased, namely, adenosine kinase, zinc finger protein X-linked and an enzyme with lyase activity. At 1 microM DCVC, two genes showed a >2-fold decrease, N-acetyltransferase 8 and complement factor H. At a lower stringency (1.5-fold change), a total of 63 probe sets were altered at 0.1 microM DCVC and 45 at 1 microM DCVC. Genes associated with stress, apoptosis, cell proliferation and repair and DCVC metabolism were altered, as were a small number of genes that did not appear to be associated with the known mode of action of DCVC. Some of these genes may serve as molecular markers of TCE exposure and effects in the human kidney.
Lock
Edward A
EA
Department of Pharmaceutical Sciences, Medical University of South Carolina, 280 Calhoun Street, PO Box 250140, Charleston, SC 29425, USA. e.lock@ljmu.ac.uk
Barth
Jeremy L
JL
Argraves
Scott W
SW
Schnellmann
Rick G
RG
D000248:Adenosine Kinase / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D000293:Adolescent; D000328:Adult; D000368:Aged; D001191:Arylamine N-Acetyltransferase / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D017242:Complement Factor H / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D003545:Cysteine / Q000031:analogs & derivatives* / Q000633:toxicity; D004247:DNA / Q000032:analysis; D004268:DNA-Binding Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004785:Environmental Pollutants / Q000633:toxicity*; D005260:Female; D015870:Gene Expression / Q000187:drug effects*; D020869:Gene Expression Profiling; D005947:Glucose / Q000378:metabolism; D005978:Glutathione / Q000031:analogs & derivatives / Q000633:toxicity; D006801:Humans; D066298:In Vitro Techniques; D007527:Isoenzymes / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D007687:Kidney Tubules, Proximal / Q000187:drug effects* / Q000473:pathology; D051741:Kruppel-Like Transcription Factors; D008190:Lyases / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D012333:RNA, Messenger / Q000378:metabolism; D014157:Transcription Factors / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D014241:Trichloroethylene / Q000378:metabolism*
D004268:DNA-Binding Proteins; D004785:Environmental Pollutants; D007527:Isoenzymes; D051741:Kruppel-Like Transcription Factors; D012333:RNA, Messenger; D014157:Transcription Factors; C062076:zinc finger protein, X-linked; D014241:Trichloroethylene; C039961:S-(1,2-dichlorovinyl)cysteine; D017242:Complement Factor H; D004247:DNA; C041464:S-(1,2-dichlorovinyl)glutathione; D001191:Arylamine N-Acetyltransferase; C089384:N-acetyltransferase 1; D000248:Adenosine Kinase; D008190:Lyases; D005978:Glutathione; D005947:Glucose; D003545:Cysteine