Prognostic significance of disseminated tumor cells as detected by quantitative real-time reverse-transcriptase polymerase chain reaction in patients with breast cancer

Clin Breast Cancer. 2006 Jun;7(2):146-52. doi: 10.3816/CBC.2006.n.024.


Background: In this study we have validated the feasibility of detecting disseminated tumor cells (DTC) by real-time reverse-transcriptase polymerase chain reaction (RT-PCR) analysis. Bone marrow samples from a large cohort of patients with breast cancer were analyzed for the presence of DTC by immunocytochemistry (ICC) or a molecular-based method.

Patients and methods: Bone marrow samples were collected from 170 patients with breast cancer with stage I-IV disease before the initiation of any local or systemic treatment. Staining for cytokeratin (CK)-positive cells was performed with the Epimet kit. Disseminated tumor cells were also quantified by measuring relative gene expression for CK19 and mammaglobin (MAM) using a quantitative RT-PCR detection method. The mean follow-up time was 30 months. Kaplan-Meier analysis was used for predicting overall survival.

Results: Despite an excellent quantitative correlation and qualitative concordance between ICC and RT-PCR, survival analysis suggested an improved prognostic significance of DTC as detected by quantitative RT-PCR. Univariate survival analysis computed a relative risk of death of 2.87 for women with ICC-positive cells in the bone marrow, as compared with those without positive cells. The relative risk for women with RT-PCR-positive bone marrow was even higher: 3.5 (CK19) and 3.39 (MAM). In multivariate analysis, bone marrow CK19 was a stronger prognostic factor than bone marrow ICC.

Conclusion: Reverse-transcriptase polymerase chain reaction-detected DTC is shown to be prognostically significant in untreated patients with breast cancer. Furthermore, it seems to be a more sensitive method for detecting DTC in bone marrow samples when compared with ICC.

Publication types

  • Controlled Clinical Trial

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Bone Marrow Neoplasms / secondary*
  • Breast Neoplasms / metabolism
  • Breast Neoplasms / pathology*
  • Feasibility Studies
  • Female
  • Humans
  • Immunohistochemistry
  • Keratins / genetics
  • Keratins / metabolism
  • Mammaglobin A
  • Middle Aged
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / metabolism
  • Neoplastic Cells, Circulating*
  • Prognosis
  • RNA, Messenger / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction*
  • Uteroglobin / genetics
  • Uteroglobin / metabolism


  • Mammaglobin A
  • Neoplasm Proteins
  • RNA, Messenger
  • SCGB2A2 protein, human
  • Keratins
  • Uteroglobin