title: Comparison of methods used to study cell death in an adherent tumoral cell line with moderate clonogenic radiosensitivity.
abstract: Our objective was to compare different methods for studying programmed cell death in adherent H460 non-small lung cancer cells of moderate clonogenic radiosensitivity. The major effect of gamma-radiation was found to be the release of cells from the substratum. The different methods gave complementary and unexpected information: a) with the TUNEL method, a few non-apoptotic cells were found in the culture medium; b) with the flow cytometry after propidium iodide labeling, some hypodiploid cells which remained attached to the substratum were apoptotic, as demonstrated by the effect of a caspase inhibitor; c) with the annexin V labeling, the detached cells were demonstrated either necrotic or very late apoptotic; d) the mitochondria transmembrane potential (deltapsim), measurements demonstrated that the mitochondria were implicated in cell death induced by gamma-radiation. These data illustrate the need to use several complementary methods in the study of apoptosis in adherent cells exposed to gamma-radiation.
authors.affiliation: INSERM U189, Faculté de Médecine Lyon-Sud, Oullins, France. broquet@lyon-sud.univ-lyon1.fr
authors.forename: P
authors.initials: P
authors.lastname: Broquet
authors.forename: C
authors.initials: C
authors.lastname: Rodriguez-Lafrasse
authors.forename: G
authors.initials: G
authors.lastname: Alphonse
authors.forename: M T
authors.initials: MT
authors.lastname: Aloy
authors.forename: P
authors.initials: P
authors.lastname: Louisot
authors.forename: R
authors.initials: R
authors.lastname: Rousson
mesh_terms: D017304:Annexin A5 / Q000494:pharmacology; D002448:Cell Adhesion; D002453:Cell Cycle / Q000528:radiation effects; D016923:Cell Death; D004171:Diploidy; D004307:Dose-Response Relationship, Radiation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005434:Flow Cytometry; D005720:Gamma Rays; D007150:Immunohistochemistry; D020287:In Situ Nick-End Labeling; D008564:Membrane Potentials; D008928:Mitochondria / Q000378:metabolism; D010718:Phosphatidylserines / Q000378:metabolism; D011419:Propidium / Q000494:pharmacology; D014407:Tumor Cells, Cultured
chemical_list: D017304:Annexin A5; D004791:Enzyme Inhibitors; D010718:Phosphatidylserines; D011419:Propidium