Pentoxifylline-Induced Apoptosis in Chronic Lymphocytic Leukemia

New Insights into Molecular Mechanism

Roberta Napolitano, Serena De Matteis, Alessandro Lucchesi, Silvia Carloni, Delia Cangini, Gerardo Musuraca, Eliana Valentina Liardo, Marianna Norata, Pier Paolo Fattori

Research output: Contribution to journalArticle

Abstract

BACKGROUND: Chronic lymphocytic leukemia (CLL) is an indolent B-lineage neoplasm, characterized by clonal expansion of CD5 positive B cells with constitutive activation of survival pathways including NF-kB. Pentoxifylline, a xanthine-derivative compound indicated for the treatment of microvascular disturbancies, has been suggested to have anti-proliferative and anti-metastatic activities in various types of cancer. In the present study we extend these data showing one of the potential molecular mechanisms through which Pentoxifylline may promote apoptosis in CLL clonal lymphocytes.

METHODS: Peripheral blood mononuclear cells were isolated from 15 CLL patients 0 RAI stage and 15 healthy volunteers and treated for 24 and 48 hours with Pentoxifylline. Apoptosis induction was evaluated through Annexin V and TUNEL assays. Mitochondrial membrane potential depolarization analysis, active Caspase-3 assay, reactive oxygen species generation and Western Blot were assessed to further investigate the alterations induced by Pentoxifylline.

RESULTS: We observed a statistically significant occurrence of apoptosis, DNA fragmentation and active Caspase-3 in lymphocytes from CLL patients compared to healthy volunteers after 48 hours of Pentoxifylline treatment. To clarify the molecular mechanism of the drug, we also evaluated the expression levels of NF-kB/p65 and its related proteins. In treated CLL cells, NF-kB/p65 was significantly decreased in comparison to normal cells, whereas we observed a less marked reduction of Bcl-2 expression. The treatment also induced a decrease of Mcl-1 in CLL cells with a greater down-regulation of the anti-apoptotic alternatively spliced isoform.

CONCLUSION: These findings showed that Pentoxifylline induced apoptosis in leukemic cells through a molecular mechanism that involves the NF-kB signaling.

Original languageEnglish
Pages (from-to)287-294
Number of pages8
JournalMini-Reviews in Medicinal Chemistry
Volume18
Issue number3
DOIs
Publication statusPublished - 2018

Fingerprint

Pentoxifylline
B-Cell Chronic Lymphocytic Leukemia
NF-kappa B
Apoptosis
Caspase 3
Healthy Volunteers
Lymphocytes
Xanthine
Mitochondrial Membrane Potential
Annexin A5
In Situ Nick-End Labeling
DNA Fragmentation
Reactive Oxygen Species
Blood Cells
Neoplasms
Protein Isoforms
B-Lymphocytes
Therapeutics
Down-Regulation
Western Blotting

Keywords

  • Antineoplastic Agents
  • Apoptosis
  • Cells, Cultured
  • Dose-Response Relationship, Drug
  • Drug Screening Assays, Antitumor
  • Humans
  • Leukemia, Lymphocytic, Chronic, B-Cell
  • Leukocytes, Mononuclear
  • Membrane Potential, Mitochondrial
  • NF-kappa B
  • Pentoxifylline
  • Proto-Oncogene Proteins c-bcl-2
  • Structure-Activity Relationship
  • Journal Article

Cite this

Pentoxifylline-Induced Apoptosis in Chronic Lymphocytic Leukemia : New Insights into Molecular Mechanism. / Napolitano, Roberta; De Matteis, Serena; Lucchesi, Alessandro; Carloni, Silvia; Cangini, Delia; Musuraca, Gerardo; Liardo, Eliana Valentina; Norata, Marianna; Fattori, Pier Paolo.

In: Mini-Reviews in Medicinal Chemistry, Vol. 18, No. 3, 2018, p. 287-294.

Research output: Contribution to journalArticle

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AU - De Matteis, Serena

AU - Lucchesi, Alessandro

AU - Carloni, Silvia

AU - Cangini, Delia

AU - Musuraca, Gerardo

AU - Liardo, Eliana Valentina

AU - Norata, Marianna

AU - Fattori, Pier Paolo

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AB - BACKGROUND: Chronic lymphocytic leukemia (CLL) is an indolent B-lineage neoplasm, characterized by clonal expansion of CD5 positive B cells with constitutive activation of survival pathways including NF-kB. Pentoxifylline, a xanthine-derivative compound indicated for the treatment of microvascular disturbancies, has been suggested to have anti-proliferative and anti-metastatic activities in various types of cancer. In the present study we extend these data showing one of the potential molecular mechanisms through which Pentoxifylline may promote apoptosis in CLL clonal lymphocytes.METHODS: Peripheral blood mononuclear cells were isolated from 15 CLL patients 0 RAI stage and 15 healthy volunteers and treated for 24 and 48 hours with Pentoxifylline. Apoptosis induction was evaluated through Annexin V and TUNEL assays. Mitochondrial membrane potential depolarization analysis, active Caspase-3 assay, reactive oxygen species generation and Western Blot were assessed to further investigate the alterations induced by Pentoxifylline.RESULTS: We observed a statistically significant occurrence of apoptosis, DNA fragmentation and active Caspase-3 in lymphocytes from CLL patients compared to healthy volunteers after 48 hours of Pentoxifylline treatment. To clarify the molecular mechanism of the drug, we also evaluated the expression levels of NF-kB/p65 and its related proteins. In treated CLL cells, NF-kB/p65 was significantly decreased in comparison to normal cells, whereas we observed a less marked reduction of Bcl-2 expression. The treatment also induced a decrease of Mcl-1 in CLL cells with a greater down-regulation of the anti-apoptotic alternatively spliced isoform.CONCLUSION: These findings showed that Pentoxifylline induced apoptosis in leukemic cells through a molecular mechanism that involves the NF-kB signaling.

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KW - Proto-Oncogene Proteins c-bcl-2

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