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

Thomas Efferth, Bernard W Futscher, Rainhardt Osieka

Research output: Contribution to journalArticle

31 Citations (Scopus)

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 drags, 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 (daunombicin, 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 drags such as cisplatin and increased cisplatin-induced DNA damage.

Original languageEnglish (US)
Pages (from-to)637-648
Number of pages12
JournalBlood Cells, Molecules, and Diseases
Volume27
Issue number3
DOIs
StatePublished - 2001

Fingerprint

Azacitidine
K562 Cells
Cytostatic Agents
Methylation
Cisplatin
Daunorubicin
Polymerase Chain Reaction
DNA Damage
Genes
CpG Islands
DNA
Vincristine
Etoposide
Drug Combinations
Leukemia, Myelogenous, Chronic, BCR-ABL Positive
Genetic Promoter Regions
Pharmaceutical Preparations
Digestion
Spectrum Analysis
Flow Cytometry

Keywords

  • Bisulfite sequencing
  • HpaII-sensitive PCR
  • PCR stop assay

ASJC Scopus subject areas

  • Molecular Biology
  • Molecular Medicine
  • Hematology

Cite this

5-Azacytidine modulates the response of sensitive and multidrug-resistant K562 leukemic cells to cytostatic drugs. / Efferth, Thomas; Futscher, Bernard W; Osieka, Rainhardt.

In: Blood Cells, Molecules, and Diseases, Vol. 27, No. 3, 2001, p. 637-648.

Research output: Contribution to journalArticle

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