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A CHO cell line resistant to deoxycholic acid
G Caderni1, E Stuart, A M Anderson
1Ludwig Institute for Cancer Research, Toronto, Ontario, Canada.
Cancer Letters
|December 1, 1987
Summary
Researchers developed a new cell line, alpha 3, exhibiting increased resistance to deoxycholic acid (DCA) and related lipids. This enhanced resistance was linked to a slight increase in specific phospholipids, suggesting potential cellular targets for bile acid action.
Area of Science:
- Cell Biology
- Biochemistry
- Toxicology
Background:
- Bile acids, such as deoxycholic acid (DCA), play crucial roles in physiological processes but can exhibit toxicity at elevated concentrations.
- Understanding cellular mechanisms of resistance to bile acids is important for various biological and medical applications.
- Chinese Hamster Ovary (CHO) cell lines are widely used models for studying cellular responses and drug resistance.
Purpose of the Study:
- To select and characterize spontaneous mutants of CHO cells with enhanced resistance to deoxycholic acid (DCA).
- To investigate the cross-resistance profile of the selected mutant cell lines against other bile acids and acidic lipids.
- To perform an initial characterization of phospholipid composition in resistant versus parental cell lines.
Main Methods:
- Selection of spontaneous deoxycholic acid (DCA)-resistant mutants from a CHO cell line (AuxBl).
- Further mutagenesis of a resistant line using ethyl methanesulfonate (EMS) to derive a highly resistant cell line (alpha 3).
- Determination of LD50 values for DCA in parental (AuxBl) and mutant (alpha 3) cell lines.
- Assessment of cross-resistance to other bile acids and acidic lipids.
- Analysis of phospholipid composition (sphingomyelin and phosphatidylcholine) using thin-layer chromatography (TLC).
Main Results:
- The mutant cell line alpha 3 demonstrated significantly higher resistance to DCA compared to the parental AuxBl cell line (LD50 of 0.66 mM vs. 0.26 mM).
- Alpha 3 cells exhibited cross-resistance to cholic acid, chenodeoxycholic acid, linoleic acid, and oleic acid.
- A statistically significant increase in the fraction of sphingomyelin and phosphatidylcholine was observed in the resistant alpha 3 cell line compared to the parental AuxBl line (54.9% vs. 51.2%).
Conclusions:
- A novel CHO cell line, alpha 3, has been successfully developed with significantly enhanced resistance to deoxycholic acid and related lipids.
- The observed cross-resistance suggests a potentially broad mechanism of lipid resistance in the alpha 3 cell line.
- Alterations in sphingomyelin and phosphatidylcholine fractions may contribute to the observed bile acid resistance, warranting further investigation into the cellular targets of bile acid action.