Foetal rat lung epithelial (FRLE) cells: alterations in cellular homeostasis and gene expression in response to etoposide, hydrogen peroxide and sodium butyrate.
Genomics technology offers a way of detecting the effects of a toxin on the expression of many genes in a single experiment. We have previously partially characterised a foetal rat lung epithelial (FRLE) cell line and shown that it is suitable for use in a pneumocytotoxicity screen. In this study, we wanted to ascertain whether we could use alterations in FRLE cell gene expression as a sensitive marker of cell stress. Sodium butyrate and etoposide were shown to arrest FRLE cell cycle at G0/G1 and G2/M phase of the cell cycle, respectively and this was associated with a decrease in the number of cells in culture. Following 24 h of culture both compounds caused a statistically significant increase in the mRNA levels of the cell cycle inhibitory protein, gadd153, whereas p21 was statistically altered by etoposide only. Hydrogen peroxide induced growth arrest at low concentrations (< or =250 microM) following 24 h of culture. We could not detect an increase in apoptosis or in the mRNA levels of the pro-apoptotic protein bax in FRLE cells following culture with hydrogen peroxide or etoposide. Thus, it was possible to correlate cellular perturbations in FRLE cells with alterations in gene expression, demonstrating that these cells are suitable for use in a toxicity screen.
Ridd
K
K
School of Biomolecular Sciences, Liverpool John Moores University, Byrom Street, Liverpool L3 3AF, UK. kr47@le.ac.uk
Alexander
D J
DJ
Reed
C J
CJ
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002087:Butyrates / Q000633:toxicity*; D022762:CCAAT-Enhancer-Binding Proteins / Q000235:genetics / Q000378:metabolism; D016466:CHO Cells; D002470:Cell Survival / Q000187:drug effects; D006224:Cricetinae; D003412:Cricetulus; D004305:Dose-Response Relationship, Drug; D004848:Epithelium / Q000187:drug effects / Q000201:enzymology / Q000473:pathology; D005047:Etoposide / Q000633:toxicity*; D005434:Flow Cytometry; D015870:Gene Expression / Q000187:drug effects*; D006706:Homeostasis / Q000187:drug effects; D006801:Humans; D006861:Hydrogen Peroxide / Q000633:toxicity*; D019169:Jurkat Cells; D008168:Lung / Q000187:drug effects* / Q000196:embryology / Q000473:pathology; D019384:Nucleic Acid Synthesis Inhibitors / Q000633:toxicity*; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D051742:Transcription Factor CHOP; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D051028:bcl-2-Associated X Protein
C494826:BAX protein, human; C494828:Bax protein, rat; D002087:Butyrates; D022762:CCAAT-Enhancer-Binding Proteins; C497249:DDIT3 protein, human; C497251:Ddit3 protein, rat; D019384:Nucleic Acid Synthesis Inhibitors; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D014157:Transcription Factors; D051028:bcl-2-Associated X Protein; D051742:Transcription Factor CHOP; D005047:Etoposide; D006861:Hydrogen Peroxide