title: Real-Time qPCR-Based Detection of Circulating Tumor Cells from Blood Samples of Adjuvant Breast Cancer Patients: A Preliminary Study.
abstract: BACKGROUND
  Circulating tumor cells (CTCs) are cells that detach from a primary tumor, circulate through the blood stream and lymphatic vessels, and are considered to be the main reason for remote metastasis. Due to their origin, tumor cells have different gene expression levels than the surrounding blood cells. Therefore, they might be detectable in blood samples from breast cancer patients by real-time quantitative polymerase chain reaction (RT-qPCR).
  
  
  METHODS
  Blood samples of healthy donors and adjuvant breast cancer patients were withdrawn and the cell fraction containing white blood cells and tumor cells was enriched by density gradient centrifugation. RNA was isolated and reverse transcribed to cDNA, which was then used in TaqMan real-time PCR against cytokeratin (CK)8, CK18 and CK19. 18S and GAPDH were used as controls.
  
  
  RESULTS
  All 3 CKs were, on average, found to be significantly higher expressed in adjuvant breast cancer samples compared to negative controls, probably due to the presence of CTCs. Unfortunately, gene expression levels could not be correlated to tumor characteristics.
  
  
  CONCLUSIONS
  RT-qPCR could make up a new approach for the detection of CTCs from blood samples of breast cancer patients, but a correlation of the PCR data to gold standard methods in CTC detection would help to further improve the informative value of the qPCR results.
authors: Department of Obstetrics and Gynecology, Ludwig Maximilians University, Munich, Germany.
authors: Ulrich
authors: U
authors: Andergassen
authors: Department of Obstetrics and Gynecology, Ludwig Maximilians University, Munich, Germany.
authors: Michael
authors: M
authors: Zebisch
authors: Department of Obstetrics and Gynecology, Ludwig Maximilians University, Munich, Germany.
authors: Alexandra C
authors: AC
authors: Kölbl
authors: Department of Obstetrics and Gynecology, Ludwig Maximilians University, Munich, Germany.
authors: Alexander
authors: A
authors: König
authors: Department of Obstetrics and Gynecology, Ludwig Maximilians University, Munich, Germany.
authors: Sabine
authors: S
authors: Heublein
authors: Department of Obstetrics and Gynecology, Ludwig Maximilians University, Munich, Germany.
authors: Lennard
authors: L
authors: Schröder
authors: Department of Obstetrics and Gynecology, Ludwig Maximilians University, Munich, Germany.
authors: Stefan
authors: S
authors: Hutter
authors: Department of Obstetrics and Gynecology, Ludwig Maximilians University, Munich, Germany.
authors: Klaus
authors: K
authors: Friese
authors: Department of Obstetrics and Gynecology, Ludwig Maximilians University, Munich, Germany.
authors: Udo
authors: U
authors: Jeschke
keywords: Adjuvant breast cancer; Circulating tumor cells; Cytokeratins; RT-qPCR