5-azacytidine modulates the response of sensitive and multidrug-resistant K562 leukemic cells to cytostatic drugs

Autoren

Thomas Efferth
Bernard W Futscher
Rainhardt Osieka

DOI

10.1006/bcmd.2001.0427

ISSN

1079-9796

Ausgabe der Veröffentlichung

3

Zeitschrift

Blood Cells, Molecules, and Diseases

Sprache

en

Paginierung

637 - 648

Datum der Veröffentlichung

2001

Status

Published

Herausgeber

Elsevier BV

Herausgeber URL

http://dx.doi.org/10.1006/bcmd.2001.0427

Datum der Datenerfassung

2023

Titel

5-Azacytidine Modulates the Response of Sensitive and Multidrug-Resistant K562 Leukemic Cells to Cytostatic Drugs

Ausgabe der Zeitschrift

27

Data source: Crossref

Abstract

In an endeavor to improve responsiveness of tumor cells to drug combination treatments, we analyzed the effect of 5-azacytidine (5AC) as a model compound for a new class of drugs, DNA-demethylating agents. We used parental K562/WT chronic myelogenous leukemia cells and a multidrug-resistant subline thereof, K562/ADM. Multidrug-resistant cells were more resistant to daunorubicin, but more sensitive to cisplatin than parental K562 cells as measured by growth inhibition and apoptosis assays. Resistance to daunorubicin can be explained by amplification of the MDR1 drug transporter gene. Cisplatin induced more DNA damage in specific genes and in the entire genome of K562/ADM cells compared to K562/WT cells using PCR stop assays and atomic absorption spectroscopy. Pretreatment with 5AC modulated the response of K562/ADM cells toward MDR-type drugs (daunorubicin, vincristine, etoposide) and reduced function and expression of MDR1 as analyzed by flow cytometry and RT-PCR. Analysis of CpG island methylation in the promotor region of the MDR1 gene by bisulfite sequencing and a methylation-sensitive HpaII-digestion/PCR approach revealed that methylation of the MDR1 promotor of K562/ADM cells was greater than in K562/WT cells. 5AC treatment completely abolished MDR1 promotor methylation. The unexpected observation that DNA demethylation by 5AC rather decreases than increases MDR1 expression in K5612/ADM cells points to still unexplored sequences in the MDR1 promotor whose transcriptional activity may be affected by the methylation status. 5AC pretreatment also modulated K562/WT and K562/ADM cells to non-MDR-type drugs such as cisplatin and increased cisplatin-induced DNA damage.

Addresses

Medizinische Klinik IV, Aachen Technical University (RWTH Aachen), Aachen, Germany. efferth@vcrp.de

Autoren

T Efferth
T Efferth
BW Futscher
R Osieka

DOI

10.1006/bcmd.2001.0427

eISSN

1096-0961

Externe Identifier

PubMed Identifier: 11482878

Open access

false

ISSN

1079-9796

Ausgabe der Veröffentlichung

3

Zeitschrift

Blood cells, molecules & diseases

Schlüsselwörter

K562 Cells
Humans
DNA Damage
Cisplatin
Azacitidine
Daunorubicin
Antineoplastic Agents
Antimetabolites, Antineoplastic
Drug Resistance, Multiple
Cell Division
Apoptosis
DNA Methylation
Gene Amplification
Drug Interactions
Genes, MDR
ATP Binding Cassette Transporter, Subfamily B, Member 1

Sprache

eng

Medium

Print

Paginierung

637 - 648

Datum der Veröffentlichung

2001

Status

Published

Datum der Datenerfassung

2001

Titel

5-Azacytidine modulates the response of sensitive and multidrug-resistant K562 leukemic cells to cytostatic drugs.

Sub types

Research Support, Non-U.S. Gov't
Journal Article

Ausgabe der Zeitschrift

27

Data source: Europe PubMed Central

Abstract

In an endeavor to improve responsiveness of tumor cells to drug combination treatments, we analyzed the effect of 5-azacytidine (5AC) as a model compound for a new class of drugs, DNA-demethylating agents. We used parental K562/WT chronic myelogenous leukemia cells and a multidrug-resistant subline thereof, K562/ADM. Multidrug-resistant cells were more resistant to daunorubicin, but more sensitive to cisplatin than parental K562 cells as measured by growth inhibition and apoptosis assays. Resistance to daunorubicin can be explained by amplification of the MDR1 drug transporter gene. Cisplatin induced more DNA damage in specific genes and in the entire genome of K562/ADM cells compared to K562/WT cells using PCR stop assays and atomic absorption spectroscopy. Pretreatment with 5AC modulated the response of K562/ADM cells toward MDR-type drugs (daunorubicin, vincristine, etoposide) and reduced function and expression of MDR1 as analyzed by flow cytometry and RT-PCR. Analysis of CpG island methylation in the promotor region of the MDR1 gene by bisulfite sequencing and a methylation-sensitive HpaII-digestion/PCR approach revealed that methylation of the MDR1 promotor of K562/ADM cells was greater than in K562/WT cells. 5AC treatment completely abolished MDR1 promotor methylation. The unexpected observation that DNA demethylation by 5AC rather decreases than increases MDR1 expression in K5612/ADM cells points to still unexplored sequences in the MDR1 promotor whose transcriptional activity may be affected by the methylation status. 5AC pretreatment also modulated K562/WT and K562/ADM cells to non-MDR-type drugs such as cisplatin and increased cisplatin-induced DNA damage.

Autoren

T Efferth
BW Futscher
R Osieka

Autoren-URL

https://www.ncbi.nlm.nih.gov/pubmed/11482878

DOI

10.1006/bcmd.2001.0427

ISSN

1079-9796

Ausgabe der Veröffentlichung

3

Zeitschrift

Blood Cells Mol Dis

Schlüsselwörter

ATP Binding Cassette Transporter, Subfamily B, Member 1
Antimetabolites, Antineoplastic
Antineoplastic Agents
Apoptosis
Azacitidine
Cell Division
Cisplatin
DNA Damage
DNA Methylation
Daunorubicin
Drug Interactions
Drug Resistance, Multiple
Gene Amplification
Genes, MDR
Humans
K562 Cells

Sprache

eng

Country

United States

Paginierung

637 - 648

PII

S1079-9796(01)90427-9

Datum der Veröffentlichung

2001

Status

Published

Datum, an dem der Datensatz öffentlich gemacht wurde

2002

Titel

5-Azacytidine modulates the response of sensitive and multidrug-resistant K562 leukemic cells to cytostatic drugs.

Sub types

Journal Article
Research Support, Non-U.S. Gov't

Ausgabe der Zeitschrift

27

Data source: PubMed

5-azacytidine modulates the response of sensitive and multidrug-resistant K562 leukemic cells to cytostatic drugs

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