Constitutively EnhancednblExpression Is Associated with the Induction of Internucleosomal DNA Cleavage by Actinomycin D

Nishida, Tsutomu; Shindo, Yutaka; Adachi, Masakazu; Naora, Honami

doi:10.1006/bbrc.1996.1017

Cited by 22 publications

(52 citation statements)

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“…6,7 The plasmacytoma HS-Sultan was cultured as described for HL-60 cells. Assessment of cell viability by trypan blue dye exclusion, and of cell morphology is described elsewhere.…”

Section: Culture and Microscopic Examination Of Human Cellsmentioning

confidence: 99%

“…Assessment of cell viability by trypan blue dye exclusion, and of cell morphology is described elsewhere. 6,8 Free cells, gently trypsinized cells or cell aggregates were prepared for Giemsa-staining by cytocentrifugation to allow detailed examination of chromatin distribution, as a result of which some cytoplasmic integrity was sacrificed. Ninety-sixwell plates were coated with poly 2-hydroxyethylmethacrylate (poly(HEMA)) (Sigma) as described.…”

Section: Culture and Microscopic Examination Of Human Cellsmentioning

confidence: 99%

“…Correction for RNA content, by hybridization with the 0.97 kb 18S ribosomal DNA fragment derived from plasmid pX1r14F, 10 is described elsewhere. 6 Isolation and agarose gel electrophoresis of genomic DNA is described elsewhere. 5 The extent of DNA fragmentation (Ͻ50 kb) was quantified by laser densitometric analysis of negatives of gel photographs.…”

Section: Culture and Microscopic Examination Of Human Cellsmentioning

confidence: 99%

“…5 More recently, we have found that apoptosis can be induced by the transcriptional inhibitor actinomycin D (Act D) in a variety of human cell lines which constitutively express the nbl gene at high levels. 6 The promyelocytic leukemia cell line HL-60 shows a particularly strong susceptibility to apoptotic induction by Act D and constitutively expresses nbl at very high levels. 6 In contrast, cell lines with low constitutive levels of nbl expression show no typical apoptotic features when treated with Act D. 6 It was therefore of interest to investigate whether apoptosis could be induced by lowering the levels of nbl expression in cells which normally express this gene at high levels.…”

Section: Introductionmentioning

confidence: 99%

“…6 The promyelocytic leukemia cell line HL-60 shows a particularly strong susceptibility to apoptotic induction by Act D and constitutively expresses nbl at very high levels. 6 In contrast, cell lines with low constitutive levels of nbl expression show no typical apoptotic features when treated with Act D. 6 It was therefore of interest to investigate whether apoptosis could be induced by lowering the levels of nbl expression in cells which normally express this gene at high levels. In this study, we examined in HL-60 cells the effects of transient expression of nbl antisense sequences and of incubation with nbl antisense phosphorothioate oligomers.…”

Section: Introductionmentioning

confidence: 99%

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Antisense sequences of the nbl gene induce apoptosis in the human promyelocytic leukemia cell line HL-60

et al. 1998

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Apoptosis is induced by the transcriptional inhibitor actinomycin D (Act D) in various cell types, particularly many leukemic cell lines such as HL-60. A common feature of these cell lines is their high constitutive expression level of the nbl gene, which was originally isolated by virtue of its abundance in a Namalwa Burkitt lymphoma cDNA library. In contrast, cell lines which constitutively express nbl at low levels appear not to undergo typical apoptotic death in response to Act D. Apoptotic induction by Act D in cells which normally express nbl at high levels was found in this study to be closely associated with a decline in nbl mRNA levels, raising the possibility that apoptosis could be induced by lowering nbl expression levels in such cells. Transient expression of nbl antisense sequences in HL-60 cells decreased cell viability, and induced typical apoptotic morphology such as cell shrinkage, chromatin condensation and nuclear fragmentation. Incubation with nbl antisense oligomers also induced similar features in HL-60 cells and in another high nbl-expressing cell line, Jurkat, but had little effect in HepG2 cells which constitutively express nbl at low levels. These findings suggest that lowering constitutively high levels of nbl expression can induce apoptosis.

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“…6,7 The plasmacytoma HS-Sultan was cultured as described for HL-60 cells. Assessment of cell viability by trypan blue dye exclusion, and of cell morphology is described elsewhere.…”

Section: Culture and Microscopic Examination Of Human Cellsmentioning

confidence: 99%

Section: Culture and Microscopic Examination Of Human Cellsmentioning

confidence: 99%

Section: Culture and Microscopic Examination Of Human Cellsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Antisense sequences of the nbl gene induce apoptosis in the human promyelocytic leukemia cell line HL-60

et al. 1998

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Lineage-related susceptibility of human hemopoietic cell lines to apoptosis

Mariani

Columbaro²,

Zauli

et al. 1999

Anat. Rec.

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Apoptosis plays a fundamental role in shaping normal hematopoiesis. We have investigated the relationship existing between susceptibility to apoptosis and lineage commitment in hemopoietic cells. The presence and degree of apoptosis were investigated in myeloid (HL-60 and K562), T (Jurkat and MOLT-4), and B (CESS and Raji) lymphoid cell lines by using a variety of techniques-transmission electron and light microscopy, flow cytometry and DNA gel electrophoresis. The major achievement of this study is that hematopoietic cells respond to different chemical (staurosporin, tiazofurin, camptothecin) and physical (hyperthermia or hypothermia) stimuli by apoptosis in a lineage-related way. Moreover, with respect to the methods used to detect apoptosis, a strong correlation was observed between the presence of the hypodiploid peak determined by flow cytometry and the DNA laddering evaluated by gel electrophoresis, but both techniques failed to demonstrate the presence of apoptosis in some cases. We conclude that cells of different hematopoietic lineages mostly show a lineage-related behaviour in their apoptotic response to different stimuli, suggesting that the lineage commitment and the stage of differentiation can confer different sensitivities to specific apoptotic stimuli. Moreover, morphological techniques still represent the most reliable approach to detect apoptosis in hemopoietic cells. Anat Rec 254:1-6, 1999. 1999 Wiley-Liss, Inc.

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Actinomycin D induces apoptosis and inhibits growth of pancreatic cancer cells

et al. 2000

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Pancreatic cancer cells are usually resistant to apoptosis induced by cytotoxic drugs, by activation of surface receptors such as Fas and TNF receptor or by serum or growth factor withdrawal. Actinomycin D (actD) is an inhibitor of RNA synthesis and acts as a potent inducer of apoptosis in several cell lines. In the present study, we investigated the effects of actD on PANC‐1 pancreatic cancer cells. ActD caused apoptosis in PANC‐1 cells in a dose‐dependent manner, as determined by cell growth assays, DNA laddering and TUNEL assays. Induction of apoptosis correlated with activation of the JNK/SAPK pathway and increased expression of Bax but not Bad or p53. PANC‐1 cells were completely resistant to Fas antibody and TNF‐α. In contrast, TRAIL decreased the growth of PANC‐1 cells by 22%. Low concentrations of actD (10 ng/ml) enhanced the cytotoxic effects of all 3 cytokines. EGF, FGF‐2 and IGF‐I did not protect PANC‐1 cells from actD‐mediated apoptosis. ActD (10 ng/ml) also inhibited the growth of CAPAN‐1 and T3M4 pancreatic cancer cells but not MiaPaCa‐2 cells. Our observations suggest that actD may act via JNK/SAPK and Bax to promote apoptosis in PANC‐1 cells and that it may inhibit the growth of other pancreatic cancer cell lines. Int. J. Cancer 86:399–407, 2000. © 2000 Wiley‐Liss, Inc.

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Constitutively EnhancednblExpression Is Associated with the Induction of Internucleosomal DNA Cleavage by Actinomycin D

Cited by 22 publications

References 0 publications

Antisense sequences of the nbl gene induce apoptosis in the human promyelocytic leukemia cell line HL-60

Antisense sequences of the nbl gene induce apoptosis in the human promyelocytic leukemia cell line HL-60

Lineage-related susceptibility of human hemopoietic cell lines to apoptosis

Actinomycin D induces apoptosis and inhibits growth of pancreatic cancer cells

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