Detection of breast cancer cell contamination in leukapheresis product by real-time quantitative polymerase chain reaction.
Identification of sensitive techniques for breast cancer cell detection might be relevant for high-dose chemotherapy programs with autologous stem cell transplantation. We investigated the feasibility of Maspin, Mammaglobin and c-ErbB-2 amplification by real-time quantitative polymerase chain reaction (RQ-PCR) for the detection of breast cancer cells in leukaphereses. Expression of the three markers was determined in primary breast cancers and cell lines. Peripheral blood (PB), bone marrow (BM), and leukapheresis samples from patients with malignancies other than breast cancer were used as controls. Sensitivity was evaluated by dilution of primary tumors and cell lines with mononuclear blood cells. We found expression of the three markers in all primary tumors and most cell lines. No blood specimen from control patients had the Maspin transcript, while only one was positive for Mammaglobin. Weak c-ErbB-2 expression was detectable in most PB, all BM and all leukapheresis samples from controls. We observed a low sensitivity of Maspin RQ-PCR and a sensitivity of Mammaglobin RQ-PCR up to one tumor cell in 10(6) mononuclear cells. One out of 18 leukaphereses from breast cancer patients screened for the presence of Mammaglobin mRNA was positive. We conclude that Mammaglobin RQ-PCR might be a useful tool for detection of leukapheresis contamination.
Leone
F
F
Ordine Mauriziano, Institute for Cancer Research and Treatment, Candiolo, Italy.
Perissinotto
E
E
Viale
A
A
Cavalloni
G
G
Taraglio
S
S
Capaldi
A
A
Piacibello
W
W
Torchio
B
B
Aglietta
M
M
D001769:Blood; D001853:Bone Marrow; D001943:Breast Neoplasms / Q000175:diagnosis* / Q000235:genetics / Q000473:pathology; D005260:Female; D005629:Frozen Sections; D016147:Genes, Tumor Suppressor; D018734:Genes, erbB-2; D006801:Humans; D007937:Leukapheresis / Q000592:standards*; D060785:Mammaglobin A; D009363:Neoplasm Proteins / Q000235:genetics; D011506:Proteins / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000592:standards*; D012680:Sensitivity and Specificity; D015842:Serine Proteinase Inhibitors / Q000235:genetics; D015843:Serpins / Q000235:genetics; D014407:Tumor Cells, Cultured; D014598:Uteroglobin / Q000235:genetics
D060785:Mammaglobin A; D009363:Neoplasm Proteins; D011506:Proteins; D012333:RNA, Messenger; C556448:SCGB2A2 protein, human; C085344:SERPIN-B5; D015842:Serine Proteinase Inhibitors; D015843:Serpins; D014598:Uteroglobin