title: Identifying cancer origin using circulating tumor cells.
abstract: Circulating tumor cells (CTCs) have become an established clinical evaluation biomarker. CTC count provides a good correlation with the prognosis of cancer patients, but has only been used with known cancer patients, and has been unable to predict the origin of the CTCs. This study demonstrates the analysis of CTCs for the identification of their primary cancer source. Twelve mL blood samples were equally dispensed on 6 CMx chips, microfluidic chips coated with an anti-EpCAM-conjugated supported lipid bilayer, for CTC capture and isolation. Captured CTCs were eluted to an immunofluorescence (IF) staining panel consisting of 6 groups of antibodies: anti-panCK, anti-CK18, anti-CK7, anti-TTF-1, anti-CK20/anti-CDX2, and anti-PSA/anti-PSMA. Cancer cell lines of lung (H1975), colorectal (DLD-1, HCT-116), and prostate (PC3, DU145, LNCaP) were selected to establish the sensitivity and specificity for distinguishing CTCs from lung, colorectal, and prostate cancer. Spiking experiments performed in 2mL of culture medium or whole blood proved the CMx platform can enumerate cancer cells of lung, colorectal, and prostate. The IF panel was tested on blood samples from lung cancer patients (n = 3), colorectal cancer patients (n = 5), prostate cancer patients (n = 5), and healthy individuals (n = 12). Peripheral blood samples found panCK(+) and CK18(+) CTCs in lung, colorectal, and prostate cancers. CTCs expressing CK7(+) or TTF-1(+), (CK20/ CDX2)(+), or (PSA/ PSMA)(+) corresponded to lung, colorectal, or prostate cancer, respectively. In conclusion, we have designed an immunofluorescence staining panel to identify CTCs in peripheral blood to correctly identify cancer cell origin.
authors.affiliation: a Graduate Institute of Life Sciences, National Defense Medical Center , Taiwan.|b Genomics Research Center, Academia Sinica , Taiwan.
authors.forename: Si-Hong
authors.initials: SH
authors.lastname: Lu
authors.affiliation: c Division of Colon and Rectal Surgery, Colorectal Section, Department of Surgery, Chang Gung Memorial Hospital, School of Medicine, Chang Gung University , Taiwan.
authors.forename: Wen-Sy
authors.initials: WS
authors.lastname: Tsai
authors.affiliation: d Division of Urology, Department of Surgery, Chang Gung Memorial Hospital, Chang Gung University , Taiwan.
authors.forename: Ying-Hsu
authors.initials: YH
authors.lastname: Chang
authors.affiliation: e Pathology and Laboratory Medicine Department, Taipei Veterans General Hospital , Taiwan.
authors.forename: Teh-Ying
authors.initials: TY
authors.lastname: Chou
authors.affiliation: d Division of Urology, Department of Surgery, Chang Gung Memorial Hospital, Chang Gung University , Taiwan.
authors.forename: See-Tong
authors.initials: ST
authors.lastname: Pang
authors.affiliation: d Division of Urology, Department of Surgery, Chang Gung Memorial Hospital, Chang Gung University , Taiwan.
authors.forename: Po-Hung
authors.initials: PH
authors.lastname: Lin
authors.affiliation: f Chest Department , Taipei Veterans General Hospital , Taiwan.
authors.forename: Chun-Ming
authors.initials: CM
authors.lastname: Tsai
authors.affiliation: a Graduate Institute of Life Sciences, National Defense Medical Center , Taiwan.|b Genomics Research Center, Academia Sinica , Taiwan.
authors.forename: Ying-Chih
authors.initials: YC
authors.lastname: Chang
mesh_terms: D014408:Biomarkers, Tumor / Q000378:metabolism*; D005260:Female; D006801:Humans; D008297:Male; D009369:Neoplasms / Q000235:genetics* / Q000378:metabolism*; D009360:Neoplastic Cells, Circulating / Q000378:metabolism*; D011379:Prognosis
chemical_list: D014408:Biomarkers, Tumor
keywords: CMx chip; Cancer cell origin; IF panel; circulating tumor cells; immunofluorescence staining; microfluidic chip; supported lipid bilayer
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