T and B leukemic cell lines exhibit different requirements for cell death: correlation between caspase activation, DFF40/DFF45 expression, DNA fragmentation and apoptosis in T cell lines but not in Burkitt's lymphoma

Abstract Introduction: Adult T-cell leukemia/lymphoma (ATL) is an aggressive peripheral T-cell neoplasm that develops after long-term infection with human T-cell leukemia virus type I (HTLV-1). Despite the recent advances in chemotherapy, allogeneic hematopoietic stem cell transplantation, and supportive care, the prognosis for patients with acute, lymphoma, or unfavorable chronic subtypes is one of the poorest among hematological malignancies. The identification of new molecular targets for ATL prevention and treatment is desired. SIRT1, a nicotinamide adenine dinucleotide (NAD+)-dependent histone/protein deacetylase, plays crucial roles in various physiological processes, including aging and apoptosis. We previously reported that ATL patients had significantly higher SIRT1 protein levels and novel small-molecule SIRT1 inhibitors are highly effective against ATL cells.1,2 Nicotinamide phosphoribosyltransferase (Nampt) also known as pre-B-cell colony-enhancing factor 1 or visfatin is a rate-limiting enzyme in NAD+ biosynthesis, and it regulates intracellular ATP levels in mammalian cells. Most cancer cells (acute myeloid leukemia [AML], acute lymphoblastic leukemia [ALL], mantle cell lymphoma [MCL], chronic lymphocytic leukemia [CLL], and T-cell lymphoma) is sensitive to low concentrations of FK866, Nampt inhibitor, as measured in cytotoxicity and clonogenic assays.3Here, we assessed how Nampt is regulated in ATL cells and leukemic cell lines. Results: We observed that ATL patients had significantly higher SIRT1 and Nampt protein levels than healthy controls. FK866 induced significant growth inhibition and apoptosis (Annexin V+ cells and TUNEL) in leukemia/lymphoma cell lines (HTLV-1-related cell lines: S1T, MT-2; Jurkat and HL60). FK866 showed potent activities with GI50values of 0.63, 3.7, 1.0, and 3,4 nM for S1T, MT-2, Jurkat, and HL60 cells, respectively. FK866 also activated caspase activity (caspase-3, 8, and 9) with DNA fragmentation. However, a caspase inhibitor did not inhibit this caspase-dependent cell death. Interestingly, FK866 increased the LC3-II-enriched protein fraction, indicating autophagosome accumulation as well as autophagy. Autophagy detection was also performed using the CytoID Autophagy detection kit. Autophagy levels are increased in the presence of STF-62247 pre-treated with bafilomycin A1, a specific inhibitor of vacuolar proton ATPase, whose inhibition is known to block the fusion of autophagosomes with lysosomes for 2 h. Thus, FK866 simultaneously caused apoptosis and autophagy. Conclusion:These results suggest that Nampt inhibitor is highly effective against ATL cells in caspase-dependent or -independent manners with autophagy, and that its clinical application might improve the prognosis of patients with this fatal disease. 1. Kozako T, Aikawa A, Shoji T, et al. High expression of the longevity gene product SIRT1 and apoptosis induction by sirtinol in adult T-cell leukemia cells. Int J Cancer. 2012;131:2044-2055. 2. Kozako T, Suzuki T, Yoshimitsu M, et al. Novel small-molecule SIRT1 inhibitors induce cell death in adult T-cell leukaemia cells. Sci Rep. 2015;5:11345. 3. Nahimana A, Attinger A, Aubry D, et al. The NAD biosynthesis inhibitor APO866 has potent antitumor activity against hematologic malignancies. Blood. 2009;113:3276-3286. Disclosures Yoshimitsu: HUYA Bioscience International: Research Funding.

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Differential activation-dependent regulation of integrin function in cultured human T-leukemic cell lines

Blood ◽

10.1182/blood.v83.4.1039.bloodjournal8341039 ◽

1994 ◽

Vol 83 (4) ◽

pp. 1039-1050 ◽

Cited By ~ 1

Author(s):

JL Mobley ◽

E Ennis ◽

Y Shimizu

Keyword(s):

T Cells ◽

T Cell ◽

Cell Lines ◽

Peripheral Blood ◽

Human Peripheral Blood ◽

Leukemic Cell ◽

Leukemic Cell Lines ◽

Beta 1 Integrin ◽

T Cell Lines ◽

Integrin Regulation

T lymphocytes isolated from human peripheral blood express beta 1 (VLA) and LFA-1 integrins, but strong binding to integrin ligands occurs only after the delivery of an activation stimulus to the T cell. To gain further insight into activation-dependent regulation of integrin function, we have analyzed integrin activity on three different T- leukemic cell lines: Jurkat, CEM, and H9. This analysis shows important mechanistic differences in integrin regulation. First, phorbol ester treatment results in increased beta 1 integrin-dependent adhesion of both Jurkat and CEM cells to fibronectin, but decreased adhesion of H9 cells. Second, certain activation stimuli that upregulate beta 1 integrin activity in peripheral T cells are nonfunctional in these T-cell lines. Third, analysis of a panel of Jurkat mutants lacking surface expression of CD2 and/or CD3 shows that CD2-mediated upregulation of beta 1 integrin activity is dependent on expression of CD3, whereas CD28-mediated upregulation is not dependent on either CD2 or CD3 expression. Fourth, all T-cell lines tested show an inability to adhere to purified ICAM-1 via LFA-1. The selective alterations in integrin regulation in these cell lines relative to peripheral blood T cells provide important insights into the intracellular processes involved in integrin activation.

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Differential activation-dependent regulation of integrin function in cultured human T-leukemic cell lines

Blood ◽

10.1182/blood.v83.4.1039.1039 ◽

1994 ◽

Vol 83 (4) ◽

pp. 1039-1050 ◽

Cited By ~ 35

Author(s):

JL Mobley ◽

E Ennis ◽

Y Shimizu

Keyword(s):

T Cells ◽

T Cell ◽

Cell Lines ◽

Peripheral Blood ◽

Human Peripheral Blood ◽

Leukemic Cell ◽

Leukemic Cell Lines ◽

Beta 1 Integrin ◽

T Cell Lines ◽

Integrin Regulation

Abstract T lymphocytes isolated from human peripheral blood express beta 1 (VLA) and LFA-1 integrins, but strong binding to integrin ligands occurs only after the delivery of an activation stimulus to the T cell. To gain further insight into activation-dependent regulation of integrin function, we have analyzed integrin activity on three different T- leukemic cell lines: Jurkat, CEM, and H9. This analysis shows important mechanistic differences in integrin regulation. First, phorbol ester treatment results in increased beta 1 integrin-dependent adhesion of both Jurkat and CEM cells to fibronectin, but decreased adhesion of H9 cells. Second, certain activation stimuli that upregulate beta 1 integrin activity in peripheral T cells are nonfunctional in these T-cell lines. Third, analysis of a panel of Jurkat mutants lacking surface expression of CD2 and/or CD3 shows that CD2-mediated upregulation of beta 1 integrin activity is dependent on expression of CD3, whereas CD28-mediated upregulation is not dependent on either CD2 or CD3 expression. Fourth, all T-cell lines tested show an inability to adhere to purified ICAM-1 via LFA-1. The selective alterations in integrin regulation in these cell lines relative to peripheral blood T cells provide important insights into the intracellular processes involved in integrin activation.

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Cannabis-induced cytotoxicity in leukemic cell lines: the role of the cannabinoid receptors and the MAPK pathway

Blood ◽

10.1182/blood-2004-03-1182 ◽

2005 ◽

Vol 105 (3) ◽

pp. 1214-1221 ◽

Cited By ~ 48

Author(s):

Thomas Powles ◽

Robert te Poele ◽

Jonathan Shamash ◽

Tracy Chaplin ◽

David Propper ◽

...

Keyword(s):

Signal Transduction ◽

Cell Death ◽

Cell Lines ◽

Leukemic Cell ◽

Cytotoxic Agents ◽

Complex Signal ◽

Signal Transduction Pathways ◽

Potent Inducer ◽

Leukemic Cell Lines

Abstract Δ9-Tetrahydrocannabinol (THC) is the active metabolite of cannabis. THC causes cell death in vitro through the activation of complex signal transduction pathways. However, the role that the cannabinoid 1 and 2 receptors (CB1-R and CB2-R) play in this process is less clear. We therefore investigated the role of the CB-Rs in mediating apoptosis in 3 leukemic cell lines and performed microarray and immunoblot analyses to establish further the mechanism of cell death. We developed a novel flow cytometric technique of measuring the expression of functional receptors and used combinations of selective CB1-R and CB2-R antagonists and agonists to determine their individual roles in this process. We have shown that THC is a potent inducer of apoptosis, even at 1 × IC50 (inhibitory concentration 50%) concentrations and as early as 6 hours after exposure to the drug. These effects were seen in leukemic cell lines (CEM, HEL-92, and HL60) as well as in peripheral blood mononuclear cells. Additionally, THC did not appear to act synergistically with cytotoxic agents such as cisplatin. One of the most intriguing findings was that THC-induced cell death was preceded by significant changes in the expression of genes involved in the mitogen-activated protein kinase (MAPK) signal transduction pathways. Both apoptosis and gene expression changes were altered independent of p53 and the CB-Rs.

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PI 3-K and T-cell activation: limitations of T-leukemic cell lines as signaling models

Trends in Immunology ◽

10.1016/s1471-4906(01)01973-1 ◽

2001 ◽

Vol 22 (9) ◽

pp. 490-496 ◽

Cited By ~ 92

Author(s):

E Astoul

Keyword(s):

T Cell ◽

Cell Lines ◽

T Cell Activation ◽

Cell Activation ◽

Leukemic Cell ◽

Leukemic Cell Lines

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Synthesis of Two Novel Homologous Polyphosphoesters Containing Aminophosphonate Units and Cytotoxicity of Some Low-Molecular and Polymeric Aminophosphonate Derivatives

Advances in Materials Science and Engineering ◽

10.1155/2018/9565401 ◽

2018 ◽

Vol 2018 ◽

pp. 1-8

Author(s):

Ivanka Kraicheva ◽

Georgi Momekov ◽

Rositsa Mihaylova ◽

Margarita Topashka-Ancheva ◽

Ivelina Tsacheva ◽

...

Keyword(s):

Cell Death ◽

Cell Lines ◽

Leukemic Cell ◽

The Novel ◽

Histological Subtype ◽

New Materials ◽

Antitumor Effects ◽

Cytotoxicity Assays ◽

Leukemic Cell Lines

Two novel polyphosphoesters containing anthracene- and furan-derived aminophosphonate moieties, namely, poly[oxyethylene(aminophosphonate-co-H-phosphonate)]s P-12 and P-13, were synthesized through an addition of poly(oxyethylene H-phosphonate) to 9-anthrylidene-furfurylamine and characterized. The novel polyphosphoester P-12 and a series of previously described anthracene-derived compounds including Schiff bases S-1 and S-2, α-aminophosphonates A-3–A-6, bis-aminophosphonate B-6, two enantiomers A-5a and A-5b, and polyphosphoesters P-8–P-11 containing aminophosphonate units were screened for antitumor activity against a panel of human leukemic cell lines, using cisplatin as a reference cytotoxic agent. As concluded from the cytotoxicity assays, both precursors S-1 and S-2 presented similar cytotoxicity profiles that are cisplatin-like only in the REH cell line. Leader compound of the α-aminophosphonates is A-4 with cell death-inducing properties fully equaling those of the referent drug in all of the screened leukemic cell lines with the only exception being the AML histological subtype HL-60. Some of the polymeric analogues elicited moderate (P-10 and P-12) to low (P-11) cytotoxic activity, whereas the polyphosphoesters P-8 and P-9 produced in vitro antitumor effects largely surpassing cisplatin’s. The compounds P-8, P-9, and A-4 could be potential new materials for anticancer therapeutic purposed.

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Induction of apoptotic cell death by direct-current treatment in human leukemic cell lines

Journal of Cancer Research and Clinical Oncology ◽

10.1007/s004320050073 ◽

1997 ◽

Vol 123 (7) ◽

pp. 370-376 ◽

Cited By ~ 2

Author(s):

Masatsugu Kurokawa ◽

Hiroshi Sakagami ◽

Fumio Kokubu ◽

Hiromichi Noda ◽

Minoru Takeda ◽

...

Keyword(s):

Cell Death ◽

Cell Lines ◽

Direct Current ◽

Apoptotic Cell ◽

Leukemic Cell ◽

Current Treatment ◽

Apoptotic Cell Death ◽

Human Leukemic Cell ◽

Leukemic Cell Lines

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Induction of apoptotic cell death by direct-current treatment in human leukemic cell lines

Journal of Cancer Research and Clinical Oncology ◽

10.1007/bf01240119 ◽

1997 ◽

Vol 123 (7) ◽

pp. 370-376 ◽

Cited By ~ 4

Author(s):

Masatsugu Kurokawa ◽

Hiroshi Sakagami ◽

Fumio Kokubu ◽

Hiromichi Noda ◽

Minoru Takeda ◽

...

Keyword(s):

Cell Death ◽

Cell Lines ◽

Direct Current ◽

Apoptotic Cell ◽

Leukemic Cell ◽

Current Treatment ◽

Apoptotic Cell Death ◽

Human Leukemic Cell ◽

Leukemic Cell Lines

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Single-cell cloning of human T-cell lines reveals clonal variation in cell death responses to chemotherapeutics

Cancer Genetics ◽

10.1016/j.cancergen.2019.06.003 ◽

2019 ◽

Vol 237 ◽

pp. 69-77 ◽

Cited By ~ 2

Author(s):

Kathleen Hanlon ◽

Alex Thompson ◽

Lorena Pantano ◽

John N. Hutchinson ◽

Arshed Al-Obeidi ◽

...

Keyword(s):

Cell Death ◽

T Cell ◽

Single Cell ◽

Cell Lines ◽

Clonal Variation ◽

Cell Cloning ◽

Human T Cell ◽

Single Cell Cloning ◽

T Cell Lines

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Overexpression of SIRT1, a Longevity Gene-Encoded Protein, and Induction of Apoptosis by Its Inhibition In Adult T-Cell Leukemia Cells

Blood ◽

10.1182/blood.v116.21.2768.2768 ◽

2010 ◽

Vol 116 (21) ◽

pp. 2768-2768

Author(s):

Tomohiro Kozako ◽

Makoto Yoshimitsu ◽

Yohann White ◽

Akiyoshi Aikawa ◽

Teruhisa Shoji ◽

...

Keyword(s):

T Cell ◽

Cell Lines ◽

Leukemic Cell ◽

Acute Type ◽

T Cell Leukemia ◽

Cell Leukemia ◽

Adult T Cell Leukemia ◽

Leukemic Cell Lines ◽

Sirt1 Inhibitor ◽

Induced Apoptosis

Abstract Abstract 2768 Adult T-cell leukemia-lymphoma (ATL) is an aggressive peripheral T-cell neoplasm developing after a long-term infection with human T-cell leukemia virus (HTLV-1), in which NF-kB is also implicated as an exacerbation factor. Despite recent progress in both chemotherapy and supportive care for hematological malignancies, the prognosis of ATL is still poor; overall survival at 3 years is only 24%. New strategies for the therapy and prophylaxis of ATL (e.g., vaccines and novel molecular target agents) are still required. SIRT1, an NAD+-dependent histone/protein deacetylase, plays a crucial role in various physiological processes, such as aging, metabolism, neurogenesis and apoptosis, due to its ability to deacetylate numerous substrates, such as histone and NF-kB. Here, we assessed how SIRT1 is regulated in primary ATL cells and leukemic cell lines. SIRT1 expression in ATL patients was significantly higher than that in healthy controls, especially in the acute type. Sirtinol, a SIRT1 inhibitor, induced significant growth inhibition or apoptosis in cells from ATL patients and leukemic cell lines, especially HTLV-1-related cell lines (S1T and MT-2). Sirtinol-induced apoptosis was mediated by activation of the caspase family, and inactivation of NF-kB, reducing IkBα phosphorylation. Interestingly, NAD+ augmented sirtinol-induced apoptosis following deacetylation of NF-kB via NAD+-dependent deacetylase. Thus, the SIRT1 inhibitor acted as a tumor suppressor, where NAD+ accelerated the SIRT1 inhibitor-induced apoptosis. These results suggest that SIRT1 is a crucial antiapoptotic molecule in ATL cells, and that SIRT1 inhibitors may be useful therapeutic agents for leukemia, especially in patients with ATL. Figure 1. SIRT1 inhibitor reduces viability of leukemic cell lines. Cell lines were incubated at 1×105 cells/mL in the presence of various concentrations of sirtinol for 72 h. Proliferation of cell lines in the absence or presence of the indicated concentrations of sirtinol was assessed by WST-8 assay. Data are means ± S.D. from 3 independent experiments. Figure 1. SIRT1 inhibitor reduces viability of leukemic cell lines. Cell lines were incubated at 1×105 cells/mL in the presence of various concentrations of sirtinol for 72 h. Proliferation of cell lines in the absence or presence of the indicated concentrations of sirtinol was assessed by WST-8 assay. Data are means ± S.D. from 3 independent experiments. Disclosures: No relevant conflicts of interest to declare.

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