Detection of bone marrow micrometastases by reverse-transcriptase polymerase chain reaction (RT-PCR) in patients with early breast cancer

Year 2008, Volume: 23 Issue: 2, 55 - 62, 01.03.2008

Neslihan Cabıoğlu Engin Okan Yıldırım Özlem Bitişik Hülya Yazıcı Mahmut Müslümanoğlu Nejat Dalay Abdullah İğci Vahit Özmen Temel Dağoğlu Mustafa Keçer

Abstract

OBJECTIVES The aim of this study was to determine the feasibility of a molecular assay by investigating cytokeratin 19 (CK-19) and DF3 gene expressions in detection of occult cancer cells in bone marrow in patients with breast cancer. METHODS Transcriptase-polymerase chain reaction (RT-PCR) assays were carried out for the detection of CK19 and DF3 mRNA transcripts in bone marrow samples of 28 clinically axillary negative patients with early breast cancer. RESULTS Median age of patients was 47.5 (range, 32-86). Due to the high false positivity rate of the CK19-RT-PCR assay, and negative findings by the DF3-RT-PCR assay in bone marrow samples of patients with chronic myelogenous leukemia as negative controls, DF3 expression was tested in further RT-PCR assays. E i g hteen patients (64%) were found to harbor DF3-positivity in bone marrow samples. No statistically significant associations could be found between DF3-positivity and other parameters including stage, tumor size (2 cm), axillary involvement, ER/PR positivity, and high histologic or nuclear grade. CONCLUSION Our results suggest that testing DF3 expression by RT-PCR technique seems to be a reliable method in detection of micrometastatic cells in bone marrow in patients with early breast cancer because of its high specificity.

Keywords

Breast Neoplasms, Reverse Transcriptase Polymerase Chain Reaction, Bone Marrow Neoplasms, Bone Marrow, Neoplasm Metastasis, Keratin-19, Gene Expression, DF3 breast-carcinoma-associated antigen

Erken evre meme kanserinde revers transkriptaz polimeraz zincir reaksiyonu (RT-PCR) tekniği ile kemik iliği mikrometastazlarının araştırılması

Abstract

AMAÇ Meme kanserinde kemik iliği mikrometastazı saptanmasında sitokeratin 19 (CK19) ve DF3 gen ekspresyonlarının araştırıldığı moleküler bir yöntemin etkinliğini değerlendirmektir. GEREÇ VE YÖNTEM Klinik olarak aksilla negatif olan 28 erken evre meme kanserli hastadan alınan kemik iliği aspiratlarında revers transkriptaz polimeraz zincir reaksiyonu (RT-PCR) tekniğiyle CK19 ve DF3 genlerine özgü iki ayrı primer kullanılarak mikrometastatik kanser hücresi araştırıldı. BULGULAR Hastaların ortanca yaşı 47,5'tir (32-86). RT-PCR çalışmalarında negatif kontrol olarak kullanılan kronik miyeloid lösemili kemik iliği örneklerinin keratin 19 ekspresyonun pozitif ve DF3 ekspresyonunun ise negatif bulunması üzerine, sonraki deneylerde DF3 primeri kullanı ldı. Yirmi sekiz hastanın 18'inde (%64) DF3 pozitif olarak saptandı. DF3 pozitifliği ile hastaların evresi, tümör büyüklüğü (< 2 cm ve > 2 cm), aksiller tutulum, ER/PR pozitifliği ve HG veya NG yüksekliği gibi parametreler arasında istatistiksel açıdan anlamlı bir fark bulunamadı. SONUÇ Erken evre meme kanserli hastalarda kemik iliğinde RT-PCR ile DF3 gen ekspresyonunun varlığının araştırılması yoluyla mikrometastaz saptanması tekniğinin, spesifikliğinin ve duyarlılığının yüksek olması nedeniyle etkin bir yöntem olduğu düşünülmektedir.

Keywords

Kemik iliği metastazları, meme kanseri, mikrometastaz, revers transkriptaz polimeraz zincir reaksiyonu.

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