title: Comparison of NMR lipid profiles in mitotic arrest and apoptosis as indicators of paclitaxel resistance in cervical cell lines.
abstract: This study aimed to characterize changes in lipid saturation using magnetic resonance spectroscopy of sensitive (HeLa) and resistant (C33A; Me180) cervical cancer cell lines following exposure to paclitaxel to explore lipid profiles as biomarkers of drug resistance. Spectra were acquired at 11.74 T. Flow cytometry, electron, and confocal microscopy assessed cellular morphology. Western blots assessed cytoplasmic phospholipase A(2) , fatty acid synthase, and acyl-CoA synthetase1 expression. After 24 h of paclitaxel exposure, >60% of cells showed mitotic arrest. At 48 h, HeLa cells showed apoptosis while C33A/Me180 cells showed normal morphology indicating resistance. MR-visible lipids increased significantly in all lines at 24 h with further increases at 48 h; resistant lines showed smaller increases than HeLa. Cytoplasmic phospholipase A(2) and fatty acid synthase levels were unchanged at 24 h and dropped at 48 h in HeLa; acyl-CoA synthetase1 was higher in Me180/C33A than in HeLa controls but did not increase significantly. The percentage of cells displaying lipid droplets increased significantly at 24 and 48 h in all lines; droplet size increased only in HeLa cells. Droplet number was >3-4× greater in apoptotic compared with mitotic-arrested cells. Apoptotic cells accumulate unsaturated fatty acids in large (relative to control) droplets; resistant lines accumulated smaller droplets with less triglycerides.
authors.affiliation: CRUK and EPSRC Cancer Imaging Centre, The Institute of Cancer Research and Royal Marsden NHS Foundation Trust, Sutton, United Kingdom.
authors.forename: Dominik
authors.initials: D
authors.lastname: Zietkowski
authors.forename: Geoffrey S
authors.initials: GS
authors.lastname: Payne
authors.forename: Eszter
authors.initials: E
authors.lastname: Nagy
authors.forename: Margaret A
authors.initials: MA
authors.lastname: Mobberley
authors.forename: Timothy A
authors.initials: TA
authors.lastname: Ryder
authors.forename: Nandita M
authors.initials: NM
authors.lastname: deSouza
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D014408:Biomarkers, Tumor / Q000032:analysis; D019008:Drug Resistance, Neoplasm; D005260:Female; D006367:HeLa Cells; D006801:Humans; D008055:Lipids / Q000032:analysis*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008938:Mitosis / Q000187:drug effects*; D017239:Paclitaxel / Q000008:administration & dosage*
chemical_list: D014408:Biomarkers, Tumor; D008055:Lipids; D017239:Paclitaxel