Cargando…

The cyclin-dependent kinase inhibitor 5, 6-dichloro-1-beta-D-ribofuranosylbenzimidazole induces nongenotoxic, DNA replication-independent apoptosis of normal and leukemic cells, regardless of their p53 status

BACKGROUND: Current chemotherapy of human cancers focuses on the DNA damage pathway to induce a p53-mediated cellular response leading to either G1 arrest or apoptosis. However, genotoxic treatments may induce mutations and translocations that result in secondary malignancies or recurrent disease. I...

Descripción completa

Detalles Bibliográficos
Autores principales: Turinetto, Valentina, Porcedda, Paola, Orlando, Luca, De Marchi, Mario, Amoroso, Antonio, Giachino, Claudia
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2743708/
https://www.ncbi.nlm.nih.gov/pubmed/19674456
http://dx.doi.org/10.1186/1471-2407-9-281
_version_ 1782171880242806784
author Turinetto, Valentina
Porcedda, Paola
Orlando, Luca
De Marchi, Mario
Amoroso, Antonio
Giachino, Claudia
author_facet Turinetto, Valentina
Porcedda, Paola
Orlando, Luca
De Marchi, Mario
Amoroso, Antonio
Giachino, Claudia
author_sort Turinetto, Valentina
collection PubMed
description BACKGROUND: Current chemotherapy of human cancers focuses on the DNA damage pathway to induce a p53-mediated cellular response leading to either G1 arrest or apoptosis. However, genotoxic treatments may induce mutations and translocations that result in secondary malignancies or recurrent disease. In addition, about 50% of human cancers are associated with mutations in the p53 gene. Nongenotoxic activation of apoptosis by targeting specific molecular pathways thus provides an attractive therapeutic approach. METHODS: Normal and leukemic cells were evaluated for their sensitivity to 5, 6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB) through cell viability and caspase activation tests. The apoptotic pathway induced by DRB was analysed by immunfluorescence and immunoblot analysis. H2AX phosphorylation and cell cycle analysis were performed to study the dependance of apoptosis on DNA damage and DNA replication, respectively. To investigate the role of p53 in DRB-induced apoptosis, specific p53 inhibitors were used. Statistical analysis on cell survival was performed with the test of independence. RESULTS: Here we report that DRB, an inhibitor of the transcriptional cyclin-dependent kinases (CDKs) 7 and 9, triggers DNA replication-independent apoptosis in normal and leukemic human cells regardless of their p53 status and without inducing DNA damage. Our data indicate that (i) in p53-competent cells, apoptosis induced by DRB relies on a cytosolic accumulation of p53 and subsequent Bax activation, (ii) in the absence of p53, it may rely on p73, and (iii) it is independent of ATM and NBS1 proteins. Notably, even apoptosis-resistant leukemic cells such as Raji were sensitive to DRB. CONCLUSION: Our results indicate that DRB represents a potentially useful cancer chemotherapeutic strategy that employs both the p53-dependent and -independent apoptotic pathways without inducing genotoxic stress, thereby decreasing the risk of secondary malignancies.
format Text
id pubmed-2743708
institution National Center for Biotechnology Information
language English
publishDate 2009
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-27437082009-09-15 The cyclin-dependent kinase inhibitor 5, 6-dichloro-1-beta-D-ribofuranosylbenzimidazole induces nongenotoxic, DNA replication-independent apoptosis of normal and leukemic cells, regardless of their p53 status Turinetto, Valentina Porcedda, Paola Orlando, Luca De Marchi, Mario Amoroso, Antonio Giachino, Claudia BMC Cancer Research Article BACKGROUND: Current chemotherapy of human cancers focuses on the DNA damage pathway to induce a p53-mediated cellular response leading to either G1 arrest or apoptosis. However, genotoxic treatments may induce mutations and translocations that result in secondary malignancies or recurrent disease. In addition, about 50% of human cancers are associated with mutations in the p53 gene. Nongenotoxic activation of apoptosis by targeting specific molecular pathways thus provides an attractive therapeutic approach. METHODS: Normal and leukemic cells were evaluated for their sensitivity to 5, 6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB) through cell viability and caspase activation tests. The apoptotic pathway induced by DRB was analysed by immunfluorescence and immunoblot analysis. H2AX phosphorylation and cell cycle analysis were performed to study the dependance of apoptosis on DNA damage and DNA replication, respectively. To investigate the role of p53 in DRB-induced apoptosis, specific p53 inhibitors were used. Statistical analysis on cell survival was performed with the test of independence. RESULTS: Here we report that DRB, an inhibitor of the transcriptional cyclin-dependent kinases (CDKs) 7 and 9, triggers DNA replication-independent apoptosis in normal and leukemic human cells regardless of their p53 status and without inducing DNA damage. Our data indicate that (i) in p53-competent cells, apoptosis induced by DRB relies on a cytosolic accumulation of p53 and subsequent Bax activation, (ii) in the absence of p53, it may rely on p73, and (iii) it is independent of ATM and NBS1 proteins. Notably, even apoptosis-resistant leukemic cells such as Raji were sensitive to DRB. CONCLUSION: Our results indicate that DRB represents a potentially useful cancer chemotherapeutic strategy that employs both the p53-dependent and -independent apoptotic pathways without inducing genotoxic stress, thereby decreasing the risk of secondary malignancies. BioMed Central 2009-08-12 /pmc/articles/PMC2743708/ /pubmed/19674456 http://dx.doi.org/10.1186/1471-2407-9-281 Text en Copyright ©2009 Turinetto et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Turinetto, Valentina
Porcedda, Paola
Orlando, Luca
De Marchi, Mario
Amoroso, Antonio
Giachino, Claudia
The cyclin-dependent kinase inhibitor 5, 6-dichloro-1-beta-D-ribofuranosylbenzimidazole induces nongenotoxic, DNA replication-independent apoptosis of normal and leukemic cells, regardless of their p53 status
title The cyclin-dependent kinase inhibitor 5, 6-dichloro-1-beta-D-ribofuranosylbenzimidazole induces nongenotoxic, DNA replication-independent apoptosis of normal and leukemic cells, regardless of their p53 status
title_full The cyclin-dependent kinase inhibitor 5, 6-dichloro-1-beta-D-ribofuranosylbenzimidazole induces nongenotoxic, DNA replication-independent apoptosis of normal and leukemic cells, regardless of their p53 status
title_fullStr The cyclin-dependent kinase inhibitor 5, 6-dichloro-1-beta-D-ribofuranosylbenzimidazole induces nongenotoxic, DNA replication-independent apoptosis of normal and leukemic cells, regardless of their p53 status
title_full_unstemmed The cyclin-dependent kinase inhibitor 5, 6-dichloro-1-beta-D-ribofuranosylbenzimidazole induces nongenotoxic, DNA replication-independent apoptosis of normal and leukemic cells, regardless of their p53 status
title_short The cyclin-dependent kinase inhibitor 5, 6-dichloro-1-beta-D-ribofuranosylbenzimidazole induces nongenotoxic, DNA replication-independent apoptosis of normal and leukemic cells, regardless of their p53 status
title_sort cyclin-dependent kinase inhibitor 5, 6-dichloro-1-beta-d-ribofuranosylbenzimidazole induces nongenotoxic, dna replication-independent apoptosis of normal and leukemic cells, regardless of their p53 status
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2743708/
https://www.ncbi.nlm.nih.gov/pubmed/19674456
http://dx.doi.org/10.1186/1471-2407-9-281
work_keys_str_mv AT turinettovalentina thecyclindependentkinaseinhibitor56dichloro1betadribofuranosylbenzimidazoleinducesnongenotoxicdnareplicationindependentapoptosisofnormalandleukemiccellsregardlessoftheirp53status
AT porceddapaola thecyclindependentkinaseinhibitor56dichloro1betadribofuranosylbenzimidazoleinducesnongenotoxicdnareplicationindependentapoptosisofnormalandleukemiccellsregardlessoftheirp53status
AT orlandoluca thecyclindependentkinaseinhibitor56dichloro1betadribofuranosylbenzimidazoleinducesnongenotoxicdnareplicationindependentapoptosisofnormalandleukemiccellsregardlessoftheirp53status
AT demarchimario thecyclindependentkinaseinhibitor56dichloro1betadribofuranosylbenzimidazoleinducesnongenotoxicdnareplicationindependentapoptosisofnormalandleukemiccellsregardlessoftheirp53status
AT amorosoantonio thecyclindependentkinaseinhibitor56dichloro1betadribofuranosylbenzimidazoleinducesnongenotoxicdnareplicationindependentapoptosisofnormalandleukemiccellsregardlessoftheirp53status
AT giachinoclaudia thecyclindependentkinaseinhibitor56dichloro1betadribofuranosylbenzimidazoleinducesnongenotoxicdnareplicationindependentapoptosisofnormalandleukemiccellsregardlessoftheirp53status
AT turinettovalentina cyclindependentkinaseinhibitor56dichloro1betadribofuranosylbenzimidazoleinducesnongenotoxicdnareplicationindependentapoptosisofnormalandleukemiccellsregardlessoftheirp53status
AT porceddapaola cyclindependentkinaseinhibitor56dichloro1betadribofuranosylbenzimidazoleinducesnongenotoxicdnareplicationindependentapoptosisofnormalandleukemiccellsregardlessoftheirp53status
AT orlandoluca cyclindependentkinaseinhibitor56dichloro1betadribofuranosylbenzimidazoleinducesnongenotoxicdnareplicationindependentapoptosisofnormalandleukemiccellsregardlessoftheirp53status
AT demarchimario cyclindependentkinaseinhibitor56dichloro1betadribofuranosylbenzimidazoleinducesnongenotoxicdnareplicationindependentapoptosisofnormalandleukemiccellsregardlessoftheirp53status
AT amorosoantonio cyclindependentkinaseinhibitor56dichloro1betadribofuranosylbenzimidazoleinducesnongenotoxicdnareplicationindependentapoptosisofnormalandleukemiccellsregardlessoftheirp53status
AT giachinoclaudia cyclindependentkinaseinhibitor56dichloro1betadribofuranosylbenzimidazoleinducesnongenotoxicdnareplicationindependentapoptosisofnormalandleukemiccellsregardlessoftheirp53status