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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Greaves, Ronda / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter


IMPACT FACTOR 2017: 3.556

CiteScore 2017: 2.34

SCImago Journal Rank (SJR) 2017: 1.114
Source Normalized Impact per Paper (SNIP) 2017: 1.188

Online
ISSN
1437-4331
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Volume 50, Issue 11

Issues

Detection of frequent ABCB1 polymorphisms by high-resolution melting curve analysis and their effect on breast carcinoma prognosis

Radka Vaclavikova / Marie Ehrlichova / Ivona Hlavata / Vaclav Pecha / Renata Kozevnikovova / Marketa Trnkova / Jan Adamek / Hege Edvardsen
  • Institute for Cancer Research, Department of Genetics, Oslo University Hospital Radiumhospitalet, Oslo, Norway
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Vessela N. Kristensen
  • Institute for Cancer Research, Department of Genetics, Oslo University Hospital Radiumhospitalet, Oslo, Norway
  • Department of Clinical Molecular Biology (EpiGen), Akerhus University Hospital, University of Oslo, Oslo, Norway
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Ivan Gut / Pavel Soucek
Published Online: 2012-06-23 | DOI: https://doi.org/10.1515/cclm-2012-0103

Abstract

Background: The ABCB1 gene encodes P-glycoprotein implicated in the development of cellular drug resistance. The aim of this study was to develop high-resolution melting (HRM) analysis for determination of ABCB1 polymorphisms and evaluate their associations with clinical data of breast carcinoma patients.

Methods: HRM analysis was designed to assess five single nucleotide polymorphisms (SNPs) in ABCB1 (rs2214102, rs1128503, rs2032582, rs2032583 and rs1045642) in genomic DNA from 103 breast carcinoma patients. Results were confirmed by direct DNA sequencing.

Results: HRM analysis revealed distinct patterns of melting curves for the respective genotypes of all followed SNPs. Sensitivity of HRM analysis compared with direct DNA sequencing was superior (97.1% vs. 93.9%). The overall accuracy of HRM was 97.6%. The coefficients of variation in replicate experiments encompassed the range 0.002%–0.038%. On the basis of the examined SNPs, one strong haplotype block containing rs2032582 and rs1128503 SNPs was identified. Significant associations of rs2032582 SNP with tumor size, negative HER-2/neu status, and family history of breast carcinoma were found. Patients carrying the ancestral homozygous genotype (GG) in rs2214102 had significantly worse progression-free survival in comparison with carriers of the non-ancestral allele (A) in the adjuvant set (p=0.005).

Conclusions: A rapid, accurate, low-cost and time-effective method for screening ABCB1 SNPs was developed. Significant associations of ABCB1 rs2032582 and rs2214102 SNPs with prognostic factors and survival of patients were found.

This article offers supplementary material which is provided at the end of the article.

Keywords: ABCB1; breast carcinoma; high resolution melting; multidrug resistance; polymorphisms

About the article

Corresponding author: Radka Vaclavikova, Toxicogenomics Unit, National Institute of Public Health, Department of Toxicology and Safety, Srobarova 48, 100 42, Prague 10, Czech Republic Phone: +420 2 67082709, Fax: +420 2 67311236


Received: 2012-02-20

Accepted: 2012-05-22

Published Online: 2012-06-23

Published in Print: 2012-11-01


Citation Information: Clinical Chemistry and Laboratory Medicine, Volume 50, Issue 11, Pages 1999–2007, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/cclm-2012-0103.

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©2012 by Walter de Gruyter Berlin Boston.Get Permission

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