title: Detection of circulating tumour cells on mRNA levels with established breast cancer cell lines.
abstract: Circulating tumour cells were detected and quantified by real-time polymerase chain reaction (PCR) in peripheral blood, based on the fact that the expression of certain genes is upregulated in tumour tissues in comparison to surrounding blood cells. Calibration curves showing gene expression as functions of the number of tumour cells within a blood sample were prepared. Blood samples were therefore spiked with cells of breast cancer cell lines, RNA was extracted, transcribed to complementary DNA (cDNA) and used in real-time PCR reaction on the Cytokeratins (CK) 8, 18 and 19. Calibration curves were generated by Microsoft™ Excel®. Relative quantification curves of gene expression in different breast cancer cell lines showed no unitary tendencies. The oscillations in the relative quantification curves of gene expression suggested an occurrence of immunological effects, leading to an apparent agglutination of added tumour cells together with the blood cells of the sample. Thus, strategies to obtain evaluable results should be considered.
authors.affiliation: Department of Obstetrics and Gynaecology, Ludwig-Maximilians University of Munich, Munich, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Zebisch
authors.affiliation: Department of Obstetrics and Gynaecology, Ludwig-Maximilians University of Munich, Munich, Germany.
authors.forename: Alexandra C
authors.initials: AC
authors.lastname: Kölbl
authors.affiliation: Department of Obstetrics and Gynaecology, Ludwig-Maximilians University of Munich, Munich, Germany.
authors.forename: Ulrich
authors.initials: U
authors.lastname: Andergassen
authors.affiliation: Department of Obstetrics and Gynaecology, Ludwig-Maximilians University of Munich, Munich, Germany.
authors.forename: Stephan
authors.initials: S
authors.lastname: Hutter
authors.affiliation: Department of Obstetrics and Gynaecology, Ludwig-Maximilians University of Munich, Munich, Germany.
authors.forename: Julia
authors.initials: J
authors.lastname: Neugebauer
authors.affiliation: Department of Obstetrics and Gynaecology, Ludwig-Maximilians University of Munich, Munich, Germany.
authors.forename: Verena
authors.initials: V
authors.lastname: Engelstädter
authors.affiliation: Department of Obstetrics and Gynaecology, Ludwig-Maximilians University of Munich, Munich, Germany.
authors.forename: Maria
authors.initials: M
authors.lastname: Günthner-Biller
authors.affiliation: Department of Obstetrics and Gynaecology, Ludwig-Maximilians University of Munich, Munich, Germany.
authors.forename: Udo
authors.initials: U
authors.lastname: Jeschke
authors.affiliation: Department of Obstetrics and Gynaecology, Ludwig-Maximilians University of Munich, Munich, Germany.
authors.forename: Klaus
authors.initials: K
authors.lastname: Friese
authors.affiliation: Department of Obstetrics and Gynaecology, Ludwig-Maximilians University of Munich, Munich, Germany.
authors.forename: Brigitte
authors.initials: B
authors.lastname: Rack
keywords: breast cancer; circulating tumour cells; detection; mRNA; quantification; real-time polymerase chain reaction