title: A New Cell Block Method for Multiple Immunohistochemical Analysis of Circulating Tumor Cells in Patients with Liver Cancer.
abstract: OBJECTIVE
  We developed a new method of detecting circulating tumor cells (CTCs) in liver cancer patients by constructing cell blocks from peripheral blood cells, including CTCs, followed by multiple immunohistochemical analysis.
  
  
  METHODS
  Cell blockswere constructed from the nucleated cell pellets of peripheral blood afterremoval of red blood cells. The blood cell blocks were obtained from 29 patients with liver cancer, and from healthy donor blood spikedwith seven cell lines. The cell blocks and corresponding tumor tissues were immunostained with antibodies to seven markers: cytokeratin (CK), epithelial cell adhesion molecule (EpCAM), epithelial membrane antigen (EMA), CK18, α-fetoprotein (AFP), Glypican 3, and HepPar1.
  
  
  RESULTS
  The average recovery rate of spiked SW620 cells from blood cell blocks was 91%. CTCs were detected in 14 out of 29 patients (48.3%); 11/23 hepatocellular carcinomas (HCC), 1/2 cholangiocarcinomas (CC), 1/1 combined HCC-CC, and 1/3 metastatic cancers. CTCs from 14 patients were positive for EpCAM (57.1%), EMA (42.9%), AFP (21.4%), CK18 (14.3%), Gypican3 and CK (7.1%, each), and HepPar1 (0%). Patients with HCC expressed EpCAM, EMA, CK18, and AFP in tissue and/or CTCs, whereas CK, HepPar1, and Glypican3 were expressed only in tissue. Only EMA was significantly associated with the expressions in CTC and tissue. CTC detection was associated with higher T stage and portal vein invasion in HCC patients.
  
  
  CONCLUSIONS
  This cell block method allows cytologic detection and multiple immunohistochemical analysis of CTCs. Our results show that tissue biomarkers of HCC may not be useful for the detection of CTC. EpCAM could be a candidate marker for CTCs in patients with HCC.
authors.affiliation: Department of Pathology, Research Institute and Hospital, National Cancer Center, Goyang, Korea.
authors.forename: Soo Jeong
authors.initials: SJ
authors.lastname: Nam
authors.affiliation: Colorectal Cancer Branch, Research Institute and Hospital, National Cancer Center, Goyang, Korea.
authors.forename: Hyun Yang
authors.initials: HY
authors.lastname: Yeo
authors.affiliation: Department of Pathology, Research Institute and Hospital, National Cancer Center, Goyang, Korea.|Colorectal Cancer Branch, Research Institute and Hospital, National Cancer Center, Goyang, Korea.
authors.forename: Hee Jin
authors.initials: HJ
authors.lastname: Chang
authors.affiliation: Center for Liver Cancer, Research Institute and Hospital, National Cancer Center, Goyang, Korea.
authors.forename: Bo Hyun
authors.initials: BH
authors.lastname: Kim
authors.affiliation: Department of Pathology, Research Institute and Hospital, National Cancer Center, Goyang, Korea.|Center for Liver Cancer, Research Institute and Hospital, National Cancer Center, Goyang, Korea.
authors.forename: Eun Kyung
authors.initials: EK
authors.lastname: Hong
authors.affiliation: Center for Liver Cancer, Research Institute and Hospital, National Cancer Center, Goyang, Korea.
authors.forename: Joong-Won
authors.initials: JW
authors.lastname: Park
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000951:Antigens, Neoplasm / Q000097:blood; D014408:Biomarkers, Tumor / Q000097:blood*; D006528:Carcinoma, Hepatocellular / Q000097:blood* / Q000235:genetics / Q000473:pathology; D019070:Cell Lineage / Q000235:genetics; D002469:Cell Separation / Q000379:methods; D000071858:Epithelial Cell Adhesion Molecule / Q000097:blood; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D053673:Glypicans / Q000097:blood; D006801:Humans; D007150:Immunohistochemistry; D053538:Keratin-18 / Q000097:blood; D008113:Liver Neoplasms / Q000097:blood* / Q000235:genetics / Q000473:pathology; D008297:Male; D008875:Middle Aged; D018396:Mucin-1 / Q000097:blood*; D009360:Neoplastic Cells, Circulating / Q000378:metabolism* / Q000473:pathology; D000509:alpha-Fetoproteins / Q000302:isolation & purification
chemical_list: D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; C499379:EPCAM protein, human; D000071858:Epithelial Cell Adhesion Molecule; C105937:GPC3 protein, human; D053673:Glypicans; C000600022:HepPar-1 protein, human; D053538:Keratin-18; D018396:Mucin-1; D000509:alpha-Fetoproteins
keywords: Circulating neoplastic cells; Hepatocellular carcinoma; Immunohistochemistry