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Published on: June 16, 2017
Profiles of gene expression changes in L5178Y mouse lymphoma cells treated with methyl methanesulfonate and sodium
Shawn D Seidel1, Barney R Sparrow, H Lynn Kan
1Toxicology and Environmental Research and Consulting, The Dow Chemical Company, 1803 Building, Midland, MI 48674, USA. sseidel2@dow.com
Abstract:
Treatment of cells with genotoxic chemicals is expected to set into motion a series of events including gene expression changes to cope with the damage. We have investigated gene expression changes in L5178Y TK(+/-) mouse lymphoma cells in culture following treatment with methyl methanesulfonate (MMS), a direct acting genotoxin, and sodium chloride (NaCl), which induces mutations in these cells through indirect mechanisms at high concentrations. The mouse lymphoma cells were treated for 4 or 24 h and the cells were harvested for RNA isolation at the end of the treatment. Analysis of the transcriptome was performed using Clontech Mouse 1.2K cDNA microarrays (1185 genes) and hybridized using 32P-labeled cDNA. The microwell methodology was used to quantify the mutagenic response. Of the genes examined, MMS altered the expression (1.5-fold or more) of only five (four at 4 h and one after 24 h treatment). NaCl altered two genes after 4 h treatment, but after 24 h it altered 19 genes (13 down- and six up-regulated). Both compounds altered the expression of several genes associated with apoptosis and NaCl altered genes involved in DNA damage/response and GTP-related proteins. This, along with other data, indicates that the widely used L5178Y TK(+/-) mouse lymphoma cells in culture are relatively recalcitrant in terms of modulating gene expression to deal with genotoxic insult.
Insights
Genotoxic chemicals like MMS and NaCl minimally altered gene expression in mouse lymphoma cells. These cells showed limited ability to modulate gene expression in response to genotoxic stress.
Area of Science:
- Toxicogenomics
- Cellular Stress Response
- Molecular Toxicology
Background:
- Genotoxic chemical exposure triggers cellular defense mechanisms, including gene expression modulation.
- L5178Y TK(+/-) mouse lymphoma cells are a standard model for genotoxicity testing.
Purpose of the Study:
- To investigate gene expression changes in L5178Y TK(+/-) cells upon exposure to methyl methanesulfonate (MMS) and sodium chloride (NaCl).
- To assess the responsiveness of these cells to genotoxic insults at the transcriptomic level.
Main Methods:
- L5178Y TK(+/-) cells were treated with MMS and NaCl for 4 or 24 hours.
- Transcriptome analysis was performed using Clontech Mouse 1.2K cDNA microarrays.
- Mutagenic response was quantified using microwell methodology.
Main Results:
- MMS (methyl methanesulfonate) affected only five genes (1.5-fold change or more).
- NaCl altered two genes at 4 hours and 19 genes at 24 hours, impacting apoptosis and DNA damage response pathways.
- Both agents showed limited modulation of gene expression in these cells.
Conclusions:
- L5178Y TK(+/-) mouse lymphoma cells exhibit a limited capacity to alter gene expression in response to genotoxic agents.
- The observed gene expression changes suggest cellular responses related to apoptosis and DNA damage pathways.
- These findings highlight the relative recalcitrance of this cell line to genotoxic stress at the gene expression level.
