Boutonnat J, Barbier M, Ronot X, Seigneurin D
Département de Biologie et de Pathologie de la Cellule, CHU A. Michallon, Grenoble, France.
Morphologie. 2000 Jun;84(265):11-5.
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.
增殖和多药耐药状态是急性髓系白血病（AML）治疗结果的关键预测指标。本研究比较了细胞周期分析（核标记）和细胞分裂分析（膜标记，PKH67），以研究用柔红霉素（DNR）和/或阿糖胞苷（Ara-C）培养的细胞的增殖情况，这两种药物是AML治疗中常用的药物。PKH是一类亲脂性荧光膜插入染料。当标记的细胞分裂时，产生的子细胞接收一半的标记，使荧光强度降低到母细胞的一半。DNR的缺点是与碘化丙啶（PI）的光谱重叠，PI是最常用的膜完整性标记物。在本研究中，使用TOTO-3评估坏死情况，TOTO-3是一种核酸标记物，在645nm以上发射荧光，可标记细胞膜破裂的细胞。比较细胞周期分析和细胞分裂分析以研究增殖情况，结果表明PKH67是分析用药物培养的细胞有丝分裂过程的首选标记物。
