JC‐1, a sensitive probe for a simultaneous detection of P‐glycoprotein activity and apoptosis in leukemic cells

Chaoui, Driss; Faussat, Anne-Marie; Majdak, Patricia; Tang, Ruoping; Perrot, Jean‐Yves; Pasco, Sabine; Klein, Christophe; Marie, Jean‐Pierre; Legrand, Ollivier

doi:10.1002/cyto.b.20100

Cited by 46 publications

(31 citation statements)

References 19 publications

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“…As mentioned, this is in contrast to effects of insulin on pancreatic b cells, as determined by LDH release, cell titer blue and FACS analysis. The possibility that the effect of insulin might involve apoptotic pathways was suggested by measurements of mitochondrial membrane potential (JC-1), Loss of mitochondrial membrane potential is indicative of apoptosis and is an early event preceding phosphatidylserine externalization and caspase activation [34][35][36]. Evidence for the possibility that insulin may indeed increase apoptosis was obtained by data showing an increase in activated caspase-3 in response to H 2 O 2 .…”

Section: Discussionmentioning

confidence: 99%

Insulin increases H2O2-induced pancreatic beta cell death

et al. 2010

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Insulin resistance results, in part, from impaired insulin signaling in insulin target tissues. Consequently, increased levels of insulin are necessary to control plasma glucose levels. The effects of elevated insulin levels on pancreatic beta (β) cell function, however, are unclear. In this study, we investigated the possibility that insulin may influence survival of pancreatic β cells. Studies were conducted on RINm, RINm5F and Min-6 pancreatic β-cells. Cell death was induced by treatment with H(2)O(2), and was estimated by measurements of LDH levels, viability assay (Cell-Titer Blue), propidium iodide staining and FACS analysis, and mitochondrial membrane potential (JC-1). In addition, levels of cleaved caspase-3 and caspase activity were determined. Treatment with H(2)O(2) increased cell death; this effect was increased by simultaneous treatment of cells with insulin. Insulin treatment alone caused a slight increase in cell death. Inhibition of caspase-3 reduced the effect of insulin to increase H(2)O(2)-induced cell death. Insulin increased ROS production by pancreatic β cells and increased the effect of H(2)O(2). These effects were increased by inhibition of IR signaling, indicative of an effect independent of the IR cascade. We conclude that elevated levels of insulin may act to exacerbate cell death induced by H(2)O(2) and, perhaps, other inducers of apoptosis.

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Section: Discussionmentioning

confidence: 99%

Insulin increases H2O2-induced pancreatic beta cell death

et al. 2010

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“…Formation of these aggregates requires normal mitochondrial membrane potential. With altered mitochondrial membrane potential following depolarization, the dye remains as a monomer in the cytoplasm and shows green fluorescence (30)(31)(32)(33). The ratio of red/green fluorescence is a function of mitochondrial membrane potential.…”

Section: Cp-31398 Reduces the Growth Of Uvb-induced Skin Tumors In Skmentioning

confidence: 99%

CP-31398 restores mutant p53 tumor suppressor function and inhibits UVB-induced skin carcinogenesis in mice

Tang

Zhu²,

Han³

et al. 2007

J. Clin. Invest.

119

100

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Mutations in the tumor suppressor p53 are detectable in over 50% of all human malignancies. Mutant p53 protein is incapable of transactivating its downstream target genes that are required for DNA repair and apoptosis. Chronic exposure to UVB induces p53 mutations and is carcinogenic in both murine and human skin. CP-31398, a styrylquinazoline compound, restores the tumor suppressor functions of mutant forms of p53 in tumor cells. However, its effectiveness in vivo remains unclear. Here, we demonstrate that CP-31398 blocked UVB-induced skin carcinogenesis and was associated with increases in p53, p21, and BclXs. CP-31398 downregulated Bcl2, proliferating nuclear cell antigen, and cyclin D1. Activation of caspase-3 and cleavage of poly (ADP-ribose) polymerase also occurred in both tumor and perilesional skin following treatment. CP-31398 induced the expression of p53-dependent target proteins, and this was followed by apoptosis in UVB-irradiated wild-type mice but not in their p53-deficient littermates. Similar effects were observed in human skin carcinoma A431 cells expressing mutant p53. In addition, CP-31398 induced mitochondrial translocation of p53, leading to changes in mitochondrial membrane permeability pore transition (MPT) and consequent cytochrome c release in these cells. Blocking MPT diminished p53 translocation and apoptosis. These studies indicate that reconstituting p53 tumor suppressor functions in vivo by small molecular weight compounds may block the pathogenesis and progression of skin cancer.

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“…The loss of mitochondrial membrane potential (ΔΨm) has been considered another hallmark of apoptosis and can be detected by JC-1 staining (19). In healthy cells, JC-1 aggregates in the mitochondria, resulting in areas of red staining known as J-aggregations.…”

Section: Resultsmentioning

confidence: 99%

Apoptosis induced by hepatitis B virus X protein in a CCL13-HBx stable cell line

et al. 2012

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Abstract. The hepatitis B virus X protein (HBx) critically modulates cell growth by inducing apoptosis or proliferation. We sought to clarify whether HBx-mediated apoptosis in a CCL13 stable cell line (Chang-HBx) with inducible HBx expression proceeds through the extrinsic (death receptormediated) and/or intrinsic (mitochondrial-mediated) pathways of apoptosis. We used western blotting, cell viability assays, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining, caspase activity assays, JC-1 staining and DNA fragmentation analysis to study the role of HBx in apoptosis. The expression of the pro-apoptotic proteins Bax and Bad and the release of cytochrome c also increased slightly upon HBx induction. JC-1 staining showed a loss of mitochondrial membrane potential upon HBx induction. Additionally, induction of HBx increased the levels of cleaved caspase-9 (intrinsic pathway), caspase-8 (extrinsic pathway) and the common effector caspase-3 as measured by western blotting. This elevation of cleaved caspase-8 or caspase-3 and caspase-9 or caspase-3 decreased in the presence of caspase-8 inhibitor Z-IETD-FMK or caspase-9 inhibitor Z-LEHD-FMK, respectively. Both inhibitors also rescued cell growth, and the caspase-8 inhibitor Z-IETD-FMK prevented apoptotic phenomena including the TUNEL signal. DNA fragmentation analysis showed that these phenomena were not detected in the presence of higher concentration of inhibitors. Our data suggest that HBx induces apoptosis through both extrinsic and intrinsic pathways.

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JC‐1, a sensitive probe for a simultaneous detection of P‐glycoprotein activity and apoptosis in leukemic cells

Cited by 46 publications

References 19 publications

Insulin increases H2O2-induced pancreatic beta cell death

Insulin increases H2O2-induced pancreatic beta cell death

CP-31398 restores mutant p53 tumor suppressor function and inhibits UVB-induced skin carcinogenesis in mice

Apoptosis induced by hepatitis B virus X protein in a CCL13-HBx stable cell line

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