A comparison of membrane properties and composition between cell lines selected and transfected for multi-drug resistance.
Cell lines selected (CHRC5) and transfected (LR-73-1A) for multi-drug resistance have total lipid compositions which are indistinguishable between resistant and parental cells. Lipid composition was evaluated by 1H NMR and the total fatty acid content by GLC. No change in surface hydrophobicity, as measured with the fluorescent probe dansyl-PE, was observed as a result of transfection of CHO cells with the mdr1 gene. However, the selected cell line, CHRC5, showed a decreased surface hydrophobicity. This decreased surface hydrophobicity was indicated by an 8 nm increase in the fluorescence emission of dansyl-PE in the CHRC5 cell line compared with the AB1. Both resistant cell lines showed an increase in the polarisation of the fluorescent probe, TMA-DPH in the plasma membranes corresponding to a 14% and a 24% change in fluorescence polarisation for the selected and transfected cell lines, respectively. This is indicative of reduced mobility of the acyl chains in the resistant cell lines. Both the CHRC5 and the transfected cell lines showed almost a 2-fold increase in the initial rate of membrane cycling. The membrane cycling could be inhibited by a known bilayer stabiliser, the N-carbobenzoxy-D-Phe-L-Phe-Gly. These results indicate that the properties of the plasma membrane from resistant cells are altered compared with their parental cell line.
Callaghan
R
R
Department of Biochemistry, McMaster University Health Sciences Centre, Hamilton, Ontario, Canada.
van Gorkom
L C
LC
Epand
R M
RM
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000818:Animals; D016466:CHO Cells / Q000502:physiology / Q000648:ultrastructure; D002462:Cell Membrane / Q000737:chemistry / Q000502:physiology*; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002849:Chromatography, Gas / Q000379:methods; D006224:Cricetinae; D003619:Dansyl Compounds; D004161:Diphenylhexatriene; D004351:Drug Resistance / Q000235:genetics / Q000502:physiology*; D005227:Fatty Acids / Q000032:analysis / Q000502:physiology; D005454:Fluorescence Polarization / Q000379:methods; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008562:Membrane Glycoproteins / Q000235:genetics / Q000502:physiology; D008563:Membrane Lipids / Q000032:analysis / Q000502:physiology*; D010714:Phosphatidylethanolamines; D000644:Quaternary Ammonium Compounds; D014162:Transfection
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D003619:Dansyl Compounds; D005227:Fatty Acids; D008562:Membrane Glycoproteins; D008563:Membrane Lipids; D010714:Phosphatidylethanolamines; D000644:Quaternary Ammonium Compounds; D004161:Diphenylhexatriene; C024284:dansyl phosphatidylethanolamine; C027917:tetramethylammonium