Single nucleotide polymorphisms modify the transporter activity of ABCG2

Kuniaki Morisaki, Robert W. Robey, C. Özvegy-Laczka, Yasumasa Honjo, Orsolya Polgar, Kenneth Steadman, B. Sarkadi, Susan E. Bates

Research output: Contribution to journalArticle

172 Citations (Scopus)

Abstract

Single nucleotide polymorphism (SNP) analyses of the ABCG2 gene have revealed three nonsynonymous SNPs resulting in the amino acid changes at V12M, Q141K and D620N. To determine whether the SNPs have an effect on drug transport, human embryonic kidney cells (HEK-293) were stably transfected with full length ABCG2 coding wild-type or SNP variants of ABCG2. In 4-day cytotoxicity assays with mitoxantrone, topotecan, SN-38 or diflomotecan, cells transfected with wild-type R482 ABCG2 showed IC50 values up to 1.2-fold to 5-fold higher than cells expressing comparable levels of Q141K ABCG2, suggesting that the Q141K SNP affects drug transport. FTC-inhibitable mitoxantrone efflux normalized to ABCG2 surface expression as assayed by the anti-ABCG2 antibody 5D3 was significantly lower in cells transfected with Q141K ABCG2 than in those transfected with wild-type R482 ABCG2 (P=0.0048). Values for V12M and D620N ABCG2 were comparable to those for wild-type R482 ABCG2. The vanadate-sensitive ATPase activity of ABCG2 was assayed in Sf9 insect cells infected with wild-type or SNP variants of ABCG2. Basal ATPase activity in cells transfected with Q141K ABCG2 was 1.8-fold lower than in cells transfected with wild-type ABCG2, but was comparable among cells expressing wild-type, V12M or D620N ABCG2. Confocal studies of ABCG2 localization revealed higher intracellular staining in the Q141K transfectants than in cells transfected with wild-type or V12M ABCG2. Decreased transport of Hoechst 33342 was observed in Sf9 cells expressing V12M ABCG2; however, this was not true in HEK-293 cells expressing V12M ABCG2. These results suggest that the Q141K SNP affects the transport efficiency of ABCG2 and may result in altered pharmacokinetics or drug-resistance profiles in clinical oncology.

Original languageEnglish
Pages (from-to)161-172
Number of pages12
JournalCancer Chemotherapy and Pharmacology
Volume56
Issue number2
DOIs
Publication statusPublished - Aug 2005

Fingerprint

Polymorphism
Single Nucleotide Polymorphism
Nucleotides
Cells
Mitoxantrone
irinotecan
Sf9 Cells
HEK293 Cells
Pharmaceutical Preparations
Topotecan
Pharmacokinetics
Oncology
Cytotoxicity
Adenosine Triphosphatases
Assays
Medical Oncology
Genes
ATP Binding Cassette Transporter, Sub-Family G, Member 2
Drug Resistance
Inhibitory Concentration 50

Keywords

  • ABCG2
  • Nucleotides
  • Polymorphism

ASJC Scopus subject areas

  • Cancer Research
  • Pharmacology
  • Oncology

Cite this

Single nucleotide polymorphisms modify the transporter activity of ABCG2. / Morisaki, Kuniaki; Robey, Robert W.; Özvegy-Laczka, C.; Honjo, Yasumasa; Polgar, Orsolya; Steadman, Kenneth; Sarkadi, B.; Bates, Susan E.

In: Cancer Chemotherapy and Pharmacology, Vol. 56, No. 2, 08.2005, p. 161-172.

Research output: Contribution to journalArticle

Morisaki, Kuniaki ; Robey, Robert W. ; Özvegy-Laczka, C. ; Honjo, Yasumasa ; Polgar, Orsolya ; Steadman, Kenneth ; Sarkadi, B. ; Bates, Susan E. / Single nucleotide polymorphisms modify the transporter activity of ABCG2. In: Cancer Chemotherapy and Pharmacology. 2005 ; Vol. 56, No. 2. pp. 161-172.
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