A CHO cell line resistant to deoxycholic acid.
Spontaneous mutants resistant to deoxycholic acid (DCA) have been selected from a CHO cell line AuxBl. One of the colonies or 'lines' selected was subsequently mutagenized by ethylmethanesulfonate (250 micrograms/ml) and a more resistant cell, named alpha 3, has been selected. When AuxBl and alpha 3 were exposed to graded concentrations of DCA for 1 h, the LD50 for the mutant strain alpha 3 was 0.66 mM compared to 0.26 mM for the parental line. AuxBl. alpha 3 showed a cross-resistance compared to AuxBl with respect to cholic acid and chenodeoxycholic acid and to other acidic lipids such as linoleic and oleic acids. As an initial step in characterization of the cell lines, the amounts of different phospholipids from whole cell extracts of the parental and mutant strains have been measured after thin-layer chromatography (TLC) separation. The results obtained demonstrated that the fraction of sphingomyelin and phosphatidylcholine over the total phospholipids was slightly increased in the resistant cell line, alpha 3, compared to the parental line, AuxBl (54.9% +/- 1.04 compared to 51.2% +/- 0.3, P = 0.01). Further characterization of cell lines resistant to bile acids could lead to the identification of the cellular target of bile acid action.
Caderni
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Ludwig Institute for Cancer Research, Toronto, Ontario, Canada.
Stuart
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E
Anderson
A M
AM
Bruce
W R
WR
D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D006224:Cricetinae; D003840:Deoxycholic Acid / Q000633:toxicity*; D010743:Phospholipids / Q000032:analysis; D013109:Sphingomyelins / Q000032:analysis
D010743:Phospholipids; D013109:Sphingomyelins; D003840:Deoxycholic Acid