Breast Cancer Research and Treatment

, Volume 138, Issue 2, pp 485–497

Disseminated tumour cells in the bone marrow in early breast cancer: morphological categories of immunocytochemically positive cells have different impact on clinical outcome

  • M. Synnestvedt
  • E. Borgen
  • E. Schlichting
  • C. B. Schirmer
  • A. Renolen
  • K. E. Giercksky
  • J. M. Nesland
  • B. Naume
Clinical trial

DOI: 10.1007/s10549-013-2439-8

Cite this article as:
Synnestvedt, M., Borgen, E., Schlichting, E. et al. Breast Cancer Res Treat (2013) 138: 485. doi:10.1007/s10549-013-2439-8

Abstract

Detection of disseminated tumour cells (DTCs) in bone marrow by immunocytochemistry (ICC) includes morphological evaluation of cytokeratin immunopositive cells. The aim of this study was to disclose the prognostic significance of different morphological categories of ICC-positive cells according to treatment status and tumour subtype. Bone marrow samples (at surgery) were analysed for the presence of cytokeratin-positive DTCs by a standard immunocytochemical method. The immunopositive cells were classified into the following categories, prior to any analysis of the association between DTCs and clinical outcome: tumour cells (TC), uninterpretable cells (UIC), hematopoietic cells (HC), and questionable HC (QHC). The analysis included 747 early breast cancer patients. Median follow-up was 84 months for relapse, and 99 months for death. The categorisation of the ICC positive cells revealed TC in 13.3 % of the patients, whereas 13.1, 17.8, and 21.4 % of the cases were positive for UIC, QHC, and HC, respectively. Analysing all patients, only TC and UIC predicted systemic relapse. Separate analysis of all patients not receiving adjuvant systemic treatment (No-Adj; n = 389) showed that only QHC were associated with reduced survival (DDFS: p = 0.008; BCSS: p = 0.004, log rank) and the presence of QHC also remained significant in multivariate analysis. Primary tumour subgroup analysis (of all patients) by hormone receptors (HR) and HER2, demonstrated that only TC/UIC had prognostic impact in the HR+/HER2− patients, whereas presence of QHC was associated with unfavourable outcome only in triple negative patients (DDFS: p = 0.004; BCSS: p = 0.024). Patients with ≥3HC had improved outcome compared to those with fewer/no HC (DDFS: p = 0.005; BCSS: p = 0.009). Hence, morphological DTC subgroups may differ in clinical significance according to primary tumour subtype and treatment status. This emphasises the importance of DTC characterisation, and separate analyses of DTC categories according to tumour subtype. Hematopoietic (“false positive”) cells might predict an immune-related favorable clinical outcome.

Keywords

Breast cancerDisseminated tumour cellsBone marrowMorphologyFalse positive cellsClassification

Abbrevations

DTCs

Disseminated tumour cells

BM

Bone marrow

FU

Follow-up

ICC

Immunocytochemical analysis

MNC

Mononuclear cells

APAAP

Alkaline phosphatase/monoclonal mouse anti-alkaline phosphatase

HC

Hematopoietic cells

TC

Tumour cells

UIC

Uninterpretable cells

pT

Histopathological primary tumor size status

pN

Histopathological lymph node status

G1, 2, 3

Histopathological grade 1–3

IHC

Immunohistochemical staining

ER

Estrogen receptor(s)

PR

Progesteron receptor(s)

HER2

Human epidermal growth factor receptor 2

TMA

Tissue microarray

FISH

Fluorescence in situ hybridization

QHC

Questionable hematopoietic cells

BCSS

Breast cancer-specific survival

DDFS

Distant disease-free survival

HR

Hormone receptor(s)

TN

Triple negative

CTCs

Circulating tumour cells

Supplementary material

10549_2013_2439_MOESM1_ESM.doc (35 kb)
Supplementary Table 1Cox multivariate analysis in patients receiving adjuvant systemic treatment (Adj). (DOC 35 kb)
10549_2013_2439_MOESM2_ESM.doc (60 kb)
Supplementary Table 2Cox multivariate analysis. Prognostic significance of DTC categories according to primary tumour subgroups. (DOC 60 kb)
10549_2013_2439_MOESM3_ESM.tif (510 kb)
Supplementary Fig. 1Survival analyses (DDFS and BCSS) for patients with (Pos) or without (Neg) QHC detected in the BM; for all No-Adj patients, and for all TN patients; only patient samples including ≥1.5x106 cells in the negative control included. P values were computed by log-rank test. (TIFF 510 kb)
10549_2013_2439_MOESM4_ESM.tif (747 kb)
Supplementary Fig. 2Distant disease-free survival among No-Adj patients with (Pos) or without (Neg) the indicated DTC subcategory (TC versus UIC versus QHC) detected in the BM; for No-Adj HR +/HER2- patients, for No-Adj HER2 + patients (HR- or +) and for No-Adj TN patients. Due to missing data in the database for a few patients, the number of patients included in the various survival analyses differ. (TIFF 747 kb)
10549_2013_2439_MOESM5_ESM.tif (721 kb)
Supplementary Fig. 3Breast cancer-specific survival among No-Adj patients with (Pos) or without (Neg) the indicated DTC subcategory (TC versus UIC versus QHC) detected in the BM; for No-Adj HR +/HER2- patients, for No-Adj HER + patients (HR- or +) and for No-Adj TN patients. Due to missing data in the database for a few patients, the number of patients included in the various survival analyses differ. (TIFF 721 kb)

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • M. Synnestvedt
    • 1
  • E. Borgen
    • 2
  • E. Schlichting
    • 3
  • C. B. Schirmer
    • 2
  • A. Renolen
    • 2
  • K. E. Giercksky
    • 4
    • 5
  • J. M. Nesland
    • 2
    • 5
  • B. Naume
    • 1
    • 6
  1. 1.Division of Surgery and Cancer Medicine, Department of OncologyOslo University Hospital, RadiumhospitaletOsloNorway
  2. 2.Department of PathologyOslo University Hospital, RadiumhospitaletOsloNorway
  3. 3.Department of SurgeryOslo University Hospital, UllevålOsloNorway
  4. 4.Department of SurgeryOslo University Hospital, RadiumhospitaletOsloNorway
  5. 5.Institute of Clinical MedicineUniversity of OsloOsloNorway
  6. 6.K.G. Jebsen Centre for Breast Cancer Research, Institute for Clinical MedicineUniversity of OsloOsloNorway