Analysis of 1H NMR-detectable mobile lipid domains for assessment of apoptosis induced by inhibitors of DNA synthesis and replication

• Published in: Cell biology international Vol. 29; no. 1; pp. 33 - 39
• Main Authors: Mikhailenko, V.M., Philchenkov, A.A., Zavelevich, M.P.
• Format: Journal Article
• Language: English
• Published: Oxford, UK Elsevier Ltd 2005; Blackwell Publishing Ltd
• Subjects: 1H NMR; Apoptosis; Apoptosis - drug effects; Cell Line, Tumor; Cell Membrane - drug effects; DNA Replication - drug effects; Etoposide - pharmacology; Humans; Hydrogen; Magnetic Resonance Spectroscopy; Membrane Lipids - physiology; Mobile lipid domains; Quercetin - pharmacology; Vidarabine - analogs & derivatives; Vidarabine - pharmacology; Mobile lipid domains; MLD; DMSO; CH 3; CH 2; 1H NMR; Apoptosis
• ISSN: 1065-6995; 1095-8355
• DOI: 10.1016/j.cellbi.2004.11.008

Cell membrane rearrangements coincident with apoptosis may contribute to the increase in the ratio of methylene (CH 2 at 1.3 ppm) to methyl (CH 3 at 0.9 ppm) resonance signal intensity as observed by proton nuclear magnetic resonance ( 1H NMR). We studied CH 2 and CH 3 resonances in cultured cell lines treated with etoposide and fludarabine or bioflavonoid quercetin. Etoposide treatment (10 μM, 18 h) resulted in 3.3 fold increase of the CH 2/CH 3 signal intensity ratio and 6.4 fold decrease in choline signal of MT4 cells. Incubation of Namalwa cells with fludarabine (3 μM, 72 h) increased the CH 2/CH 3 signal intensity ratio by 2.4 fold and choline resonance intensity was unchanged. Quercetin treatment (30 μM, 1.5 month) increased CH 2/CH 3 ratio by 2.1 fold. Necrotic cell death upon ethanol (20%) or DMSO (30%) treatment did not change the CH 2/CH 3 signal intensity ratio. 1H NMR-based study of mobile lipid domains is sensitive for detection of early engagement into apoptosis.