title: Circulating biomarkers in hepatocellular carcinoma.
abstract: OBJECTIVE
  Our aims are to determine levels of circulating cellular and protein biomarkers in hepatocellular carcinoma (HCC) patients and to analyse any relationships with clinical parameters.
  
  
  METHODS
  Fifty-four consenting patients were recruited. Circulating tumour cells (CTCs) were enumerated (by CellSearch) and characterised via filtration [by isolation by size of epithelial tumour cells (ISET)] with downstream immunohistochemistry (IHC). Glypican-3 (GPC3) expression in tumour biopsies and CTCs (by IHC) was compared, and levels of circulating caspase-cleaved and full-length cytokeratin 18 (CK18, measured using M30 and M65 ELISAs) were examined as a putative prognostic factor and marker of tumour burden.
  
  
  RESULTS
  CTCs were identified in 14 out of 50 (28%) patients by CellSearch and in 19 out of 19 (100%) patients by ISET. The presence of GPC3-positive CTCs by ISET was 100% concordant with the presence of GPC3-positive cells in the original tumour (n = 5). No statistically significant correlations were observed between CTC number and clinical characteristics, although trends were noted between CTC subtypes, Child-Pugh score and tumour node metastasis stage. Serum M30 and M65 levels (as continuous variables) significantly correlated with overall survival (OS) in a univariate analysis (p = 0.003 and p < 0.001, respectively); M65 levels remained statistically significant in a multivariate analysis (p = 0.029).
  
  
  CONCLUSIONS
  This is the first study to detect GPC3-positive CTCs in HCC, important for drug development with this target. The significant association of circulating CK18 with OS in HCC further exemplifies the utility of circulating biomarkers in cancer.
authors.affiliation: Clinical and Experimental Pharmacology Group, Cancer Research UK Manchester Institute, University of Manchester, Manchester, UK.
authors.forename: Karen L
authors.initials: KL
authors.lastname: Morris
authors.forename: Jonathan D
authors.initials: JD
authors.lastname: Tugwood
authors.forename: Leila
authors.initials: L
authors.lastname: Khoja
authors.forename: Matthew
authors.initials: M
authors.lastname: Lancashire
authors.forename: Robert
authors.initials: R
authors.lastname: Sloane
authors.forename: Debbie
authors.initials: D
authors.lastname: Burt
authors.forename: Patrick
authors.initials: P
authors.lastname: Shenjere
authors.forename: Cong
authors.initials: C
authors.lastname: Zhou
authors.forename: Clare
authors.initials: C
authors.lastname: Hodgson
authors.forename: Toshihiko
authors.initials: T
authors.lastname: Ohtomo
authors.forename: Atsuhiko
authors.initials: A
authors.lastname: Katoh
authors.forename: Takahiro
authors.initials: T
authors.lastname: Ishiguro
authors.forename: Juan W
authors.initials: JW
authors.lastname: Valle
authors.forename: Caroline
authors.initials: C
authors.lastname: Dive
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D014408:Biomarkers, Tumor / Q000097:blood*; D006528:Carcinoma, Hepatocellular / Q000097:blood / Q000175:diagnosis*; D002469:Cell Separation; D005260:Female; D005500:Follow-Up Studies; D053673:Glypicans / Q000097:blood; D006801:Humans; D007124:Immunoenzyme Techniques; D053538:Keratin-18 / Q000097:blood; D008113:Liver Neoplasms / Q000097:blood / Q000175:diagnosis*; D008297:Male; D008875:Middle Aged; D009367:Neoplasm Staging; D009360:Neoplastic Cells, Circulating / Q000473:pathology*; D011379:Prognosis
chemical_list: D014408:Biomarkers, Tumor; C105937:GPC3 protein, human; D053673:Glypicans; D053538:Keratin-18