Abstract
9672 Background: Early hematogenous spread of occult tumor cells is a crucial event in tumor progression and formation of distant metastases. Several markers mainly cytokeratins have been validated to date using immunohistochemistry (IHC) or more sensitive reverse transcriptase polymerase chain reaction (RT-PCR). The role of Cytokeratin 19 (CK-19) remains controversial regarding it's sensitivity and specificity. In contrast Mammaglobin was reported as a breast specific marker with very high specificity. In this feasibility study we validated expression of CK19, Mammaglobin A and B in the bone marrow samples of patients with breast cancer. Methods: We obtained bone marrow aspirates of 48 patients with stage I (38%), II (58%) and III (4%) breast cancer who underwent either immediate complete resection of the tumor or neoadjuvant therapy with subsequent curative surgery. mRNA was isolated using QIAamp RNA blood mini kit (Qiagen ®). Subsequently two-step nested RT-PCR for the expression of CK-19, Mammaglobin A and B was performed. Results: CK-19 was detected in all 48 samples. Mammaglobin A was detected in samples from 6 (12,5%) out of 48 patients. Two samples (4%) were positive for Mammaglobin B. Only one patient had both Mammaglobin A and B transcripts in the bone marrow. With a median follow-up of 20 (4–33) month we observed only 2 recurrences, both in patients without occult tumor cells in the bone marrow. Conclusions: CK-19 showed neither diagnostic nor prognostic value as a mRNA marker in the RT-PCR detection of occult tumor cells in the bone marrow. Mammaglobin A may be a useful tool for detection of occult breast cancer cells in the bone marrow but it's clinical and prognostic relevance should be further investigated. Mammaglobin B showed lower sensitivity and should be further validated. Acknowledgement: Supported by a grant IGA MZ CR NC/6672–3 and MSMT CR CEZ: J13/98 1111 0000 4. No significant financial relationships to disclose.
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