title: Technical limits of comparison of step-sectioning,immunohistochemistry and RT-PCR on breast cancer sentinel nodes: a study on methacarn-fixed tissue.
abstract: The optimal pathological assessment of sentinel nodes (SLNs) in breast cancer is a matter of debate. Currently, multilevel histological evaluation and immunohistochemistry (IHC) are recommended, but alternative RT-PCR procedures have been developed. To assess the reliability of these different procedures, we devised a step-sectioning protocol at 100 micron-intervals of 74 SLNs using methacarn fixation. mRNA was extracted from sections collected from levels 4 to 5. Mammaglobin, CEA and CK19 were used for RT-PCR. mRNA extraction was successful in 69 SLNs. Of these, 7 showed macrometastases (>2mm), 2 showed micrometastases (<2 mm) and 7 showed isolated tumour cells (ITC) by IHC. RT-PCR was positive for the three markers in 6 of 7 macrometastases and in 1 of 2 micrometastases. In the 2 RT-PCR negative cases, metastases were detected only on sections distant from those analysed by RT-PCR. CEA and/or CK19 were positive by RT-PCR in 3 of 7 ITC and in 23 morphologically negative SLNs. In conclusion, the main goal of our study was to show that the use of alternate sections of the same sample for different procedures is the key reason for the discrepancies between molecular and morphological analyses of SLN. We believe that only prospective studies with quantitative mRNA analysis of specific metastatic markers on the whole lymph node can elucidate the utility of molecular assessments of SLN.
authors.affiliation: Department of Biomedical Sciences and Human Oncology, University of Turin, Turin, Italy.
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Daniele
authors.forename: Laura
authors.initials: L
authors.lastname: Annaratone
authors.forename: Elena
authors.initials: E
authors.lastname: Allia
authors.forename: Sara
authors.initials: S
authors.lastname: Mariani
authors.forename: Enrico
authors.initials: E
authors.lastname: Armando
authors.forename: Martino
authors.initials: M
authors.lastname: Bosco
authors.forename: Luigia
authors.initials: L
authors.lastname: Macrì
authors.forename: Paola
authors.initials: P
authors.lastname: Cassoni
authors.forename: Giuseppe
authors.initials: G
authors.lastname: D'Armento
authors.forename: Gianni
authors.initials: G
authors.lastname: Bussolati
authors.forename: Gabor
authors.initials: G
authors.lastname: Cserni
authors.forename: Anna
authors.initials: A
authors.lastname: Sapino
mesh_terms: D019342:Acetic Acid / Q000737:chemistry*; D000368:Aged; D001943:Breast Neoplasms / Q000378:metabolism*; D002725:Chloroform / Q000737:chemistry*; D017931:DNA Primers / Q000737:chemistry; D005260:Female; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D008495:Medical Oncology / Q000379:methods; D000432:Methanol / Q000737:chemistry*; D008875:Middle Aged; D009362:Neoplasm Metastasis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D021701:Sentinel Lymph Node Biopsy / Q000379:methods*
chemical_list: D017931:DNA Primers; C046544:methacarn; D002725:Chloroform; D019342:Acetic Acid; D000432:Methanol