Nucleus labeling or membrane labeling for studying the proliferation of drug treated cells?
Proliferation and multidrug resistance status are key predictors of therapeutic outcome in acute myeloid leukemia (AML). This study compared cell cycle analysis (nuclear labeling) with cell division analysis (membrane labeling, PKH67) for studying the proliferation of cells cultured with daunorubicin (DNR) and/or Cytarabine (Ara-C), drugs commonly used in AML treatment. PKHs are a family of lipophilic, fluorescent membrane intercalating dyes. When labeled cells divide, the resulting daughter cells receive half the label, reducing fluorescence intensity to one-half that of the parent cells. DNR has the disadvantage of overlapping the spectrum of propidium iodide (PI), which is the most commonly used marker of membrane integrity. In this study, necrosis was evaluated using TOTO-3, a marker of nucleic acids emitting fluorescence above 645 nm and which incorporates cells with disrupted membranes. Comparison of cell cycle analysis with cell division for studying proliferation revealed that PKH67 was a marker of choice for analyzing the mitotic process in cells cultured with drugs.
Boutonnat
J
J
Département de Biologie et de Pathologie de la Cellule, CHU A. Michallon, Grenoble, France. Jean.boutonnat@ujf-grenoble.fr
Barbier
M
M
Ronot
X
X
Seigneurin
D
D
D000208:Acute Disease; D000970:Antineoplastic Agents / Q000494:pharmacology*; D002453:Cell Cycle / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002462:Cell Membrane / Q000737:chemistry / Q000648:ultrastructure*; D002463:Cell Membrane Permeability / Q000187:drug effects*; D002467:Cell Nucleus / Q000737:chemistry / Q000648:ultrastructure*; D003561:Cytarabine / Q000494:pharmacology*; D004273:DNA, Neoplasm / Q000032:analysis; D003630:Daunorubicin / Q000494:pharmacology*; D018432:Drug Resistance, Multiple; D019008:Drug Resistance, Neoplasm; D005434:Flow Cytometry; D005456:Fluorescent Dyes / Q000032:analysis*; D006801:Humans; D020014:K562 Cells / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D007951:Leukemia, Myeloid / Q000188:drug therapy / Q000473:pathology*; D011419:Propidium / Q000032:analysis*
D000970:Antineoplastic Agents; D004273:DNA, Neoplasm; D005456:Fluorescent Dyes; D003561:Cytarabine; D011419:Propidium; D003630:Daunorubicin