Advances in Clinical and Experimental Medicine

Title abbreviation: Adv Clin Exp Med
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Advances in Clinical and Experimental Medicine

2009, vol. 18, nr 1, January-February, p. 19–24

Publication type: original article

Language: English

Genotyping FOXA1 G559A Polymorphism in Breast Cancer Archival Samples Using the SNaPshot Technique

Genotypowanie polimorfizmu FOXA1 G559A w archiwalnych preparatach raka gruczołu piersiowego techniką SNaPshot

Małgorzata Jaremko1,2,, Anna Sadakierska−chudy1,, Tadeusz Dobosz1,

1 Department of Forensic Medicine, Molecular Techniques Unit, Wroclaw Medical University, Poland

2 Mount Sinai School of Medicine, Department of Genetic and Genomics Sciences, NY, USA

Abstract

Background. FOXA1, a forkhead family transcription factor, is expressed in breast cancer cells and plays an essential role in the regulation of approximately 50% of estrogen receptor alpha (ERα)−dependent genes. Recent studies of expression signatures of breast cancer types have indicated its potential use as a prognostic factor in luminal type A breast cancer (BC).
Objectives. To establish the frequencies of FOXA1 G559A polymorphisms in a Polish population of breast cancer patients diagnosed with early breast carcinoma and to test whether the SNaPshot methodology is useful for genotyping DNA extracted from formalin−fixed paraffin−embedded tissues (FFPETs).
Material and Methods. DNA was extracted from 70 FFPET blocks and FOXA1 G559A polymorphism was successfully genotyped in 51 DNA samples using SNaPshot technology (Applied Biosystems).
Results. It is shown that the SNapShot technology based on the single−base primer extension reaction is suitable for genotyping DNA recovered from archived breast cancer tissues. The rate of successful amplifications was 94.4%. No genotypes deviated from Hardy−Weinberg equilibrium. The observed distribution of the FOXA1 G559A genotypes for this cohort of BC patients was GG 84%, GA 14%, and AA 2%. Conclusion. SNaPshot is a credible method for the analysis of DNA extracted from FFPETs and it can be used in large−scale FOXA1 G559A analysis. Further study of this polymorphism and analysis of patients’ case histories may provide better insight into its utility as a putative marker of breast cancer prognosis.

Streszczenie

Wprowadzenie. FOXA1 to czynnik transkrypcyjny z rodziny Forkhead, ekspresjonowany w komórkach raka piersi, odgrywający kluczową rolę w regulacji około 50% genów zależnych od receptora estrogenowego alfa (ERα). Ostatnie badania niektórych typów raka gruczołu piersiowego wykazały, że FOXA1 może być potencjalnym czynnikiem prognostycznym w luminalnym raku typu A.
Cel pracy. Ustalenie częstości polimorfizmu FOXA1 G559A w populacji polskiej pacjentek ze zdiagnozowanym wczesnym rakiem piersi oraz określenie przydatności metody SNaPshot w genotypowaniu DNA pochodzącego z utrwalonego materiału, przechowywanego w postaci bloczków parafinowych.
Materiał i metody. DNA izolowano z 70 bloczków parafinowych, a genotypowanie udało się przeprowadzić z sukcesem tylko w 51 próbkach z użyciem metody SNaPshot (Applied Biosystems).
Wyniki. Wykazano, że technika SNaPshot oparta na reakcji wydłużania starterów o jedną parę zasad pozwala na genotypowanie DNA pochodzącego z archiwalnych tkanek raka piersi. Wskaźnik udanych amplifikacji wyniósł 94,4%. Stwierdzone genotypy były ilościowo zgodne z przewidywaniami wynikającymi z prawa Hardy’ego−Weinberga. Obserwowany rozkład genotypów w badanej grupie pacjentek wynosił: GG 84%, GA 14%, AA 2%.
Wnioski. Metoda SNaPshot okazała się niezawodną metodą analizy DNA z tkanek utrwalanych formaliną i przechowywanych w formie bloczków parafinowych i może być wykorzystana do analizy polimorfizmu FOXA1 G559Aw większej grupie pacjentów. Dalsze badania polimorfizmu, w połączeniu z analizą historii choroby, są konieczne do oceny jego przydatności jako markera prognostycznego w raku piersi.

Key words

FOXA1, luminal breast cancer, SNaPshot

Słowa kluczowe

FOXA1, luminalny rak piersi, SNaPshot

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