Multicenter study using paraffin-embedded tumor tissue testing PITX2 DNA methylation as a marker for outcome prediction in tamoxifen-treated, node-negative breast cancer patients.
OBJECTIVE
We recently reported DNA methylation of the paired-like homeodomain transcription factor 2 (PITX2) gene to be strongly correlated with increased risk of recurrence in node-negative, hormone receptor-positive, tamoxifen-treated breast cancer patients using fresh frozen specimens. Aims of the present study were to establish determination of PITX2 methylation for routine analysis in formalin-fixed paraffin-embedded (FFPE) breast cancer tissue and to test PITX2 DNA methylation as a biomarker for outcome prediction in an independent patient cohort.


METHODS
Real-time polymerase chain reaction (PCR) technology was validated for FFPE tissue by comparing methylation measurements in FFPE specimens with those in fresh frozen specimens from the same tumor. The impact of PITX2 methylation on time to distant metastasis was then evaluated in FFPE specimens from hormone receptor-positive, node-negative breast cancer patients (n = 399, adjuvant tamoxifen monotherapy).


RESULTS
Reproducibility of the PCR assay in replicate measurements (r(s) > or = 0.95; n = 150) and concordant measurements between fresh frozen and FFPE tissues (r(s) = 0.81; n = 89) were demonstrated. In a multivariate model, PITX2 methylation added significant information (hazard ratio = 2.35; 95% CI, 1.20 to 4.60) to established prognostic factors (tumor size, grade, and age).


CONCLUSIONS
PITX2 methylation can be reliably assessed by real-time PCR technology in FFPE tissue. Together with our earlier studies, we have accumulated substantial evidence that PITX2 methylation analysis holds promise as a practical assay for routine clinical use to predict outcome in node-negative, tamoxifen-treated breast cancer, which might allow, based on future validation studies, the identification of low-risk patients who may be treated by tamoxifen alone.
Harbeck
Nadia
N
Department of Obstetrics and Gynecology and Pathology, Technical University of Munich, Munich, Germany. nadia.harbeck@lrz.tu-muenchen.de
Nimmrich
Inko
I
Hartmann
Arndt
A
Ross
Jeffrey S
JS
Cufer
Tanja
T
Grützmann
Robert
R
Kristiansen
Glen
G
Paradiso
Angelo
A
Hartmann
Oliver
O
Margossian
Astrid
A
Martens
John
J
Schwope
Ina
I
Lukas
Antje
A
Müller
Volkmar
V
Milde-Langosch
Karin
K
Nährig
Jörg
J
Foekens
John
J
Maier
Sabine
S
Schmitt
Manfred
M
Lesche
Ralf
R
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001118:Argentina; D014408:Biomarkers, Tumor / Q000235:genetics*; D001943:Breast Neoplasms / Q000188:drug therapy* / Q000235:genetics* / Q000473:pathology; D019175:DNA Methylation; D005060:Europe; D005260:Female; D005629:Frozen Sections; D015972:Gene Expression Regulation, Neoplastic; D018398:Homeodomain Proteins / Q000235:genetics*; D006801:Humans; D008198:Lymph Nodes / Q000473:pathology; D008875:Middle Aged; D009362:Neoplasm Metastasis; D009518:New York; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction; D011237:Predictive Value of Tests; D015203:Reproducibility of Results; D020845:Selective Estrogen Receptor Modulators / Q000627:therapeutic use*; D013629:Tamoxifen / Q000627:therapeutic use*; D013997:Time Factors; D014157:Transcription Factors / Q000235:genetics*; D016896:Treatment Outcome; D000097577:Homeobox Protein PITX2
D014408:Biomarkers, Tumor; D018398:Homeodomain Proteins; D020845:Selective Estrogen Receptor Modulators; D013629:Tamoxifen; D014157:Transcription Factors; D000097577:Homeobox Protein PITX2