title: Circulating tumor and cancer stem cells in hepatitis C virus-associated liver disease.
abstract: OBJECTIVE
  To assess the role of circulating tumor cells (CTCs) and cancer stem cells (CSCs) in hepatitis C virus (HCV)-associated liver disease.
  
  
  METHODS
  Blood and/or tissue samples were obtained from HCV (genotype 4)-associated hepatocellular carcinoma patients (HCC; n = 120), chronic hepatitis C patients (CH; n = 30) and 33 normal control subjects (n = 33). Serum levels of alpha-fetoprotein (AFP), alkaline phosphatase, and alanine and aspartate aminotransferases were measured. Cytokeratin 19 (CK19) monoclonal antibody was used to enumerate CTCs, and CD133 and CD90 were used to enumerate CSCs by flow cytometry. The expression levels of the CSCs markers (CD133 and CD90) as well as telomerase, melanoma antigen encoding gene 1 (MAGE1) and MAGE3 were assessed by RT-PCR and quantitative real-time polymerase chain reactions. The number of CTCs and/or the expression levels of CK19, CD133, telomerase, MAGE1 and MAGE3 were correlated to the standard clinicopathologic prognostic factors and disease progression.
  
  
  RESULTS
  Levels of AFP, alkaline phosphatase and aspartate aminotransferase were significantly different among the HCC, CH and control groups (P < 0.001), whereas alanine aminotransferase differed significantly between patient (HCC and CH) and control groups (P < 0.001). At the specified cutoff values determine by flow cytometry, CK19 (49.8), CD90 (400) and CD133 (73) were significantly higher in the blood of HCC patients compared to those in the CH and control groups (P < 0.001). On the other hand, CD133 at a 69.5 cutoff was significantly higher in the CH compared to the control group (P ≤ 0.001). Telomerase, MAGE1 and MAGE3 RNA were expressed in 55.71%, 60.00% and 62.86% of the HCC patients, respectively, but were not detected in patients in the CH or control groups, which were statistically significant (Ps < 0.001). The expression levels of telomerase, CD90, MAGE3, CD133 and CK19 were all significantly associated with high tumor grade and advanced stage in HCC patients (all Ps < 0.05).
  
  
  CONCLUSIONS
  CTC counts and AFP, CK19, telomerase, and MAGE1/MAGE3 expression predict disease progression in patients with HCV, whereas telomerase, MAGE3, CD90, CD133 and CK19 are prognostic markers in HCC.
authors.affiliation: Abeer A Bahnassy, Molecular Pathology Unit, Pathology Department, National Cancer Institute, Cairo University, Cairo 11796, Egypt.
authors.forename: Abeer A
authors.initials: AA
authors.lastname: Bahnassy
authors.affiliation: Abeer A Bahnassy, Molecular Pathology Unit, Pathology Department, National Cancer Institute, Cairo University, Cairo 11796, Egypt.
authors.forename: Abdel-Rahman N
authors.initials: AR
authors.lastname: Zekri
authors.affiliation: Abeer A Bahnassy, Molecular Pathology Unit, Pathology Department, National Cancer Institute, Cairo University, Cairo 11796, Egypt.
authors.forename: Ahmed
authors.initials: A
authors.lastname: El-Bastawisy
authors.affiliation: Abeer A Bahnassy, Molecular Pathology Unit, Pathology Department, National Cancer Institute, Cairo University, Cairo 11796, Egypt.
authors.forename: Amal
authors.initials: A
authors.lastname: Fawzy
authors.affiliation: Abeer A Bahnassy, Molecular Pathology Unit, Pathology Department, National Cancer Institute, Cairo University, Cairo 11796, Egypt.
authors.forename: Marwa
authors.initials: M
authors.lastname: Shetta
authors.affiliation: Abeer A Bahnassy, Molecular Pathology Unit, Pathology Department, National Cancer Institute, Cairo University, Cairo 11796, Egypt.
authors.forename: Nehal
authors.initials: N
authors.lastname: Hussein
authors.affiliation: Abeer A Bahnassy, Molecular Pathology Unit, Pathology Department, National Cancer Institute, Cairo University, Cairo 11796, Egypt.
authors.forename: Dalia
authors.initials: D
authors.lastname: Omran
authors.affiliation: Abeer A Bahnassy, Molecular Pathology Unit, Pathology Department, National Cancer Institute, Cairo University, Cairo 11796, Egypt.
authors.forename: Abdallah A S
authors.initials: AA
authors.lastname: Ahmed
authors.affiliation: Abeer A Bahnassy, Molecular Pathology Unit, Pathology Department, National Cancer Institute, Cairo University, Cairo 11796, Egypt.
authors.forename: Samir S
authors.initials: SS
authors.lastname: El-Labbody
mesh_terms: D000328:Adult; D000368:Aged; D014408:Biomarkers, Tumor / Q000097:blood / Q000235:genetics; D006528:Carcinoma, Hepatocellular / Q000097:blood / Q000235:genetics / Q000473:pathology* / Q000821:virology; D016022:Case-Control Studies; D002469:Cell Separation / Q000379:methods; D018450:Disease Progression; D005260:Female; D005434:Flow Cytometry; D005838:Genotype; D016174:Hepacivirus / Q000235:genetics; D019698:Hepatitis C, Chronic / Q000150:complications* / Q000175:diagnosis / Q000821:virology; D006801:Humans; D008113:Liver Neoplasms / Q000097:blood / Q000235:genetics / Q000473:pathology* / Q000821:virology; D008297:Male; D008875:Middle Aged; D060787:Neoplasm Grading; D009367:Neoplasm Staging; D009360:Neoplastic Cells, Circulating / Q000378:metabolism / Q000473:pathology*; D014411:Neoplastic Stem Cells / Q000378:metabolism / Q000473:pathology*; D011237:Predictive Value of Tests; D011446:Prospective Studies; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D055815:Young Adult
chemical_list: D014408:Biomarkers, Tumor
keywords: Cancer stem cells; Circulating tumor cells; Hepatitis C virus genotype-4; Hepatocellular carcinoma