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Consequences of combining siRNA-mediated DNA methyltransferase 1 depletion with 5-aza-2′-deoxycytidine in human leukemic KG1 cells
- Source :
- Oncotarget, Oncotarget, Impact journals, 2015, 6 (17), pp.15265-15282. ⟨10.18632/oncotarget.3317⟩, Scopus-Elsevier, Oncotarget, 2015, 6 (17), pp.15265-15282. ⟨10.18632/oncotarget.3317⟩, Europe PubMed Central
- Publication Year :
- 2015
- Publisher :
- HAL CCSD, 2015.
-
Abstract
- // Stephane Vispe 1 , Arthur Deroide 1 , Emeline Davoine 1 , Cecile Desjobert 1 , Fabrice Lestienne 2 , Lucie Fournier 1 , Natacha Novosad 1 , Sophie Breand 3 , Jerome Besse 3 , Florence Busato 4 , Jorg Tost 4 , Luc De Vries 2 , Didier Cussac 2 , Joelle Riond 1 , Paola B. Arimondo 1 1 Unite de Service et de Recherche n°3388 CNRS-Pierre Fabre, ETaC Epigenetic Targeting of Cancer, CRDPF, Toulouse, France 2 Molecular and Cellular Biology Department, Centre de Recherche Pierre Fabre, Castres, France 3 Informatique de Recherche (Bioinformatics and Statistics), Centre de Recherche Pierre Fabre, Castres, France 4 Laboratory for Epigenetics and Environment, Centre National de Genotypage, CEA-Institut de Genomique, Evry, France Correspondence to: Stephane Vispe, e-mail: stephane.vispe@pierre-fabre.com Paola B. Arimondo, e-mail: paola.arimondo@etac.cnrs.fr Keywords: leukemia, DNA methylation, DNMT, 5-aza-2’-deoxycytidine, DNA damage Received: December 28, 2014 Accepted: February 08, 2015 Published: March 20, 2015 ABSTRACT 5-azacytidine and 5-aza-2′-deoxycytidine are clinically used to treat patients with blood neoplasia. Their antileukemic property is mediated by the trapping and the subsequent degradation of a family of proteins, the DNA methyltransferases (DNMT1, DNMT3A, and DNMT3B) leading to DNA demethylation, tumor suppressor gene re-expression and DNA damage. Here we studied the respective role of each DNMT in the human leukemia KG1 cell line using a RNA interference approach. In addition we addressed the role of DNA damage formation in DNA demethylation by 5-aza-2′-deoxycytidine. Our data show that DNMT1 is the main DNMT involved in DNA methylation maintenance in KG1 cells and in mediating DNA damage formation upon exposure to 5-aza-2′-deoxycytidine. Moreover, KG1 cells express the DNMT1 protein at a level above the one required to ensure DNA methylation maintenance, and we identified a threshold for DNMT1 depletion that needs to be exceeded to achieve DNA demethylation. Most interestingly, by combining DNMT1 siRNA and treatment with low dose of 5-aza-2′-deoxycytidine, it is possible to uncouple DNA damage formation from DNA demethylation. This work strongly suggests that a direct pharmacological inhibition of DNMT1, unlike the use of 5-aza-2′-deoxycytidine, should lead to tumor suppressor gene hypomethylation and re-expression without inducing major DNA damage in leukemia.
- Subjects :
- [SDV]Life Sciences [q-bio]
DNMT
MESH: G2 Phase Cell Cycle Checkpoints
DNA Methyltransferase 3A
Histones
MESH: DNA Methylation
MESH: RNA, Small Interfering
DNA (Cytosine-5-)-Methyltransferases
Cancer epigenetics
Phosphorylation
RNA, Small Interfering
Promoter Regions, Genetic
MESH: CpG Islands
MESH: Histones
DNA methylation
biology
leukemia
Nuclear Proteins
3. Good health
DNA-Binding Proteins
G2 Phase Cell Cycle Checkpoints
Oncology
embryonic structures
Azacitidine
MESH: Tumor Protein p73
RNA Interference
Research Paper
5-aza-2′-deoxycytidine
DNA (Cytosine-5-)-Methyltransferase 1
MESH: DNA (Cytosine-5-)-Methyltransferases
MESH: Cell Line, Tumor
DNA repair
DNA damage
MESH: RNA Interference
MESH: Azacitidine
Decitabine
DNA methyltransferase
5-aza-2’-deoxycytidine
Cell Line, Tumor
MESH: Cell Proliferation
MESH: Leukemia
MESH: Promoter Regions, Genetic
Humans
[CHIM]Chemical Sciences
MESH: Tumor Suppressor Proteins
Epigenetics
5-aza-2'-deoxycytidine
Cell Proliferation
MESH: DNA (Cytosine-5-)-Methyltransferase 1
MESH: DNA Damage
MESH: Humans
MESH: Phosphorylation
MESH: Decitabine
Tumor Suppressor Proteins
Tumor Protein p73
Molecular biology
Proliferating cell nuclear antigen
DNA demethylation
biology.protein
Cancer research
CpG Islands
MESH: Nuclear Proteins
MESH: DNA-Binding Proteins
Subjects
Details
- Language :
- English
- ISSN :
- 19492553
- Database :
- OpenAIRE
- Journal :
- Oncotarget, Oncotarget, Impact journals, 2015, 6 (17), pp.15265-15282. ⟨10.18632/oncotarget.3317⟩, Scopus-Elsevier, Oncotarget, 2015, 6 (17), pp.15265-15282. ⟨10.18632/oncotarget.3317⟩, Europe PubMed Central
- Accession number :
- edsair.doi.dedup.....66560f2157e2f74b389fb7912920127c