Comparative study of immunocytochemical staining versus Giemsa stain for detecting dysmegakaryopoiesis in myelodysplastic syndromes (MDS)

Kawaguchi, Masato; Nehashi, Y; Aizawa, Shin; Toyama, Keisuke

doi:10.1111/j.1600-0609.1990.tb00356.x

Cited by 7 publications

References 15 publications

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Myelodysplastic Syndromes

2014

Morphology of Blood Disorders

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FAS gene, a direct target for NFκB transcription factor, is repressed in solid tumors including colon carcinomas. We have previously reported that, while overexpressed in low risk myelodysplastic syndromes (MDS), the Fas death receptor becomes undetectable on CD34 + progenitors when the disease progresses to secondary acute myeloid leukaemia (sAML). This study aimed at investigating the interplay between NFκB and Fas during MDS progression. We first observed that Fas was inducible by TNFα in the HL60 cell line. In these cells, p65/RelA bound to the chromatin at FAS promoter, while inhibition of NFκB pathway by IKKα inhibitor BAY11-7082 or lentiviral expression of a non-degradable mutant of IκBα, IκSR blocked the expression of Fas. By contrast, TNFα failed to induce Fas expression in the colon carcinoma cell line SW480, due to hypermethylation of the FAS promoter. In vitro use of azacitidine rescued p65/RelA binding on FAS promoter and, subsequently Fas expression in SW480 cells. We also show that, inhibition of NFκB pathway decreased the expression of Fas in MDS CD45 lo CD34 + bone marrow cells. However, despite of the nuclear expression of p65/RelA, Fas was often low on CD45 lo CD34 + AML cells. TNFα failed to stimulate its expression, while azacitidine efficiently rescued p65/RelA binding and Fas reexpression. Our data suggest that DNA methylation at NFκB sites is responsible for FAS gene silencing. Rescuing FAS gene expression by azacitidine could contribute to slacken the progression to leukaemia.

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Myelodysplastic Syndromes

2014

Morphology of Blood Disorders

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Comparison of the DNA content of megakaryocytes identified immunologically with that identified morphologically

Kobayashi

Takahashi²,

Chikayama³

et al. 1997

Histochemistry and Cell Biology

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We devised a new microfluorometric method for determining the ploidy of megakaryocytes identified immunologically in bone marrow smears. The smears were immunostained by incubation with mouse monoclonal anti-glycoproteins (GP) IIb antibodies, followed by fluorescein isothiocyanate-conjugated goat antimouse IgG antibodies. They were then stained with 4',6-diamidino-2-phenylindole (DAPI). Megakaryocytes were identified by their GPIIb immunofluorescence using a microfluorometer and, after the filters were changed, their DNA content was assayed by measuring the intensity of DAPI fluorescence. This intensity was shown to be proportional to the DNA content when the aperture of the objective lens was reduced. We compared these results with those obtained when megakaryocytes were identified morphologically, using DAPI staining after Wright-Giemsa destaining. In all 12 normal controls, the ploidy peaks were shown to be 16N by both methods, and the mean ploidy detected by the immunological method was only reduced 0.961 times relative to the estimate from the morphological method. In contrast, in eight myelodysplastic syndrome (MDS) patients, the ploidy peaks were either 8N or 4N and the mean was reduced by 0.906 times (P = 0.018). Thus we could immunologically identify small megakaryocytes which we could not identify morphologically. Therefore, this method is useful for measuring megakaryocytic ploidy, especially in the pathological megakaryocytes of MDS patients.

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Myelodysplastic syndromes

Noël

Solberg

1992

Critical Reviews in Oncology/Hematology

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Comparative study of immunocytochemical staining versus Giemsa stain for detecting dysmegakaryopoiesis in myelodysplastic syndromes (MDS)

Cited by 7 publications

References 15 publications

Myelodysplastic Syndromes

Myelodysplastic Syndromes

Comparison of the DNA content of megakaryocytes identified immunologically with that identified morphologically

Myelodysplastic syndromes

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