Detection of circulating tumor cells is improved by drug-induced antigen up-regulation: preclinical and clinical studies.
(51)Cr-prelabelled colon cancer cells (simulating 'circulating tumor cells', CTCs) were added to human peripheral blood and exposed to staurosporine (ST) to increase carcinoembryonic antigen (CEA) expression. CTCs were captured with immunomagnetic beads coated with Ber-EP4 monoclonal antibody, recognizing the common epithelial antigen present in the majority of cancer cells of epithelial origin, with capture efficiency of more than 80%. Moreover, ST treatment increased CEA expression without compromising Ber-EP4 capture efficiency. In a pilot clinical study on 37 patients, CTCs were captured using Ber-EP4 beads, and recognized by RT-PCR set for CEA or cytokeratin-19 (CK) mRNA detection. The results showed that: (a) the percentage of CEA-positive CTCs (CTC(CEA), 54.1%) was lower than that of CK-positive CTCs (CTC(CK), 70.3%); (b) in vitro ST treatment converted a significant number of CTC(CEA)-negative into CTC(CEA)-positive cases. Therefore, immunomagnetic capture combined with exposure to ST provides a feasible and sensitive technique for the detection of functionally-active CTCs responsive to ST-mediated CEA up-regulation.
Laboratory of Molecular Oncology, Istituto Dermopatico dell'Immacolata-IRCCS, via dei Monti di Creta 104, 00167 Rome, Italy.
Laura
L
Bonmassar
Emanuela
E
Fossile
Alessandro
A
Scoppola
Grazia
G
Graziani
Salvatore P
SP
Prete
Vincenzo
V
Formica
Daniela
D
Cappelletti
Liana
L
De Vecchis
Anna
A
Cardillo
Francesco
F
Concolino
Stefania
S
D'Atri
Alessandra
A
Balduzzi
Francesco
F
Torino
Patrizia
P
Caporaso
Jack W
JW
Greiner
Enzo
E
Bonmassar
Mario
M
Roselli
Angelo
A
Aquino
D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism; D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology; D014408:Biomarkers, Tumor / Q000032:analysis*; D002272:Carcinoembryonic Antigen / Q000235:genetics* / Q000378:metabolism; D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000097:blood* / Q000276:immunology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D005472:Fluorouracil / Q000494:pharmacology; D006801:Humans; D015151:Immunoblotting; D018189:Immunomagnetic Separation; D053539:Keratin-19 / Q000235:genetics* / Q000378:metabolism; D008297:Male; D051379:Mice; D008875:Middle Aged; D009362:Neoplasm Metastasis; D009360:Neoplastic Cells, Circulating / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019311:Staurosporine / Q000494:pharmacology*; D015854:Up-Regulation
D000911:Antibodies, Monoclonal; D000964:Antimetabolites, Antineoplastic; C117805:Ber-EP4 monoclonal antibody; D014408:Biomarkers, Tumor; D002272:Carcinoembryonic Antigen; D004791:Enzyme Inhibitors; D053539:Keratin-19; D012333:RNA, Messenger; D019311:Staurosporine; D005472:Fluorouracil