Utilization of extracellular lipids by HT29/219 cancer cells in culture.
Uptake and incorporation of long-chain fatty acids were studied in a human colorectal cancer cell line (HT29/219) grown in culture medium supplemented with either fetal calf serum (FCS) or horse serum (HS). The cells were grown for 120 h with no change of medium; the two major cellular lipid classes, the phospholipids and the triacylglycerols, were analyzed at regular time-points. We observed significant changes in the concentration of most fatty acids throughout culture, and differences in their composition when different sera were used to supplement the medium. Minimal levels of free fatty acids were found in the cells, indicating a very small "free fatty acid pool". A major difference between the cells grown in media supplemented with different sera was the changes observed in concentrations of cellular polyunsaturated fatty acids during growth. In cells grown with FCS (in which 20:4n-6 is present), the levels of this acid in the phospholipid and triacylglycerol fractions declined rapidly during cell growth, suggesting further metabolism. In cells grown in medium supplemented with HS, 18:2n-6 was the major polyunsaturated acid present. There was clear evidence that this acid accumulated in the cellular triacylglycerol and phospholipid fractions. Furthermore, its concentration did not decline during growth in culture, suggesting minimal conversion to other polyunsaturated n-6 acids. Our results suggest that fatty acids from additional sources in the medium, for example triacylglycerols and phospholipids associated with the lipoproteins, are taken up by the cells. There is also indication of cellular fatty acid synthesis, particularly of monounsaturated and saturated acids during the culture period.(ABSTRACT TRUNCATED AT 250 WORDS)
Department of Biochemical Medicine, Ninewells Hospital and Medical School, Dundee, Scotland.
S
S
Pazouki
J D
JD
Baty
H M
HM
Wallace
C S
CS
Coleman
D016718:Arachidonic Acid / Q000378:metabolism; D002455:Cell Division; D015179:Colorectal Neoplasms / Q000378:metabolism*; D003470:Culture Media; D005227:Fatty Acids / Q000378:metabolism*; D005231:Fatty Acids, Unsaturated / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D019787:Linoleic Acid; D008041:Linoleic Acids / Q000378:metabolism; D008042:Linolenic Acids / Q000378:metabolism; D019301:Oleic Acid; D009829:Oleic Acids / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism*; D014280:Triglycerides / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D017962:alpha-Linolenic Acid
D003470:Culture Media; D005227:Fatty Acids; D005231:Fatty Acids, Unsaturated; D008041:Linoleic Acids; D008042:Linolenic Acids; D009829:Oleic Acids; D010743:Phospholipids; D014280:Triglycerides; D017962:alpha-Linolenic Acid; D016718:Arachidonic Acid; D019301:Oleic Acid; D019787:Linoleic Acid