title: A blood-based three-gene signature for the non-invasive detection of early human hepatocellular carcinoma.
abstract: BACKGROUND
  Identifying early stages of disease in high-risk individuals for the development of hepatocellular carcinoma (HCC) would greatly improve the clinical outcomes of these individuals. The aim of this study was to develop a blood-based gene set that could identify early-stage HCC.
  
  
  METHODS
  Comprehensive gene expression profiling of purified RNA of peripheral blood mononuclear cells (PBMC) was performed using microarrays. Gene signatures were developed through bioinformatics-driven approaches and their diagnostic value was evaluated by custom-designed, quantitative, multiplex polymerase chain reaction (PCR) assays.
  
  
  RESULTS
  Bioinformatics-driven analysis of microarray data derived from PBMC RNA samples of patients with HCC (N=10), pancreatic cancer (N=3), gastric cancer (N=3) and 10 normal individuals identified six genes that were differentially expressed in HCC. Subsequent multiplex-PCR validation and univariate analyses performed with an independent cohort of 114 HCC patients, 48 normal individuals and 14 patients with chronic hepatitis B (CHB) validated that three genes, namely Chemokine (C-X-C motif) receptor 2 (CXCR2), C-C chemokine receptor type 2 (CCR2) and E1A-Binding Protein P400 (EP400), were able to identify HCC individually with accuracies of 82.4%, 78.4% and 65%, respectively. In combination, these three genes gave an area under the curve (AUC) of 0.96 (95% confidence interval (CI), 0.93-0.99) using multivariate logistic regression and yielded a sensitivity of 93% and a specificity of 89%. When these three genes were used in combination with alpha-fetoprotein (AFP) to predict HCC, the accuracy of predicting HCC improved slightly with an AUC of 0.99 (95% CI, 0.98-1.0), sensitivity of 93% and specificity of 95%.
  
  
  CONCLUSIONS
  CXCR2, CCR2 and EP400 can provide a promising non-invasive multiplex PCR diagnostic assay to monitor high-risk individuals for the development of HCC.
authors: Department of Hepatobiliary Oncology, Sun Yat-Sen University Cancer Center, Guangzhou 510060, PR China.
authors: Ming
authors: M
authors: Shi
authors: Department of Hepatobiliary Oncology, Sun Yat-Sen University Cancer Center, Guangzhou 510060, PR China.
authors: Min-Shan
authors: MS
authors: Chen
authors: Division of Cellular and Molecular Research, Humphrey Oei Institute of Cancer Research, National Cancer Centre, Singapore 169610, Singapore.
authors: Karthik
authors: K
authors: Sekar
authors: Department of Gastroenterology and Hepatology, Singapore General Hospital, Singapore 169608, Singapore.
authors: Chee-Kiat
authors: CK
authors: Tan
authors: Division of Surgical Oncology, National Cancer Centre, Singapore 169610, Singapore.
authors: London Lucien
authors: LL
authors: Ooi
authors: Division of Cellular and Molecular Research, Humphrey Oei Institute of Cancer Research, National Cancer Centre, Singapore 169610, Singapore; Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597, Singapore; Institute of Molecular and Cell Biology, A(∗)STAR, Biopolis Drive Proteos, Singapore 138673, Singapore; Program in Cancer and Stem Cell Biology, Duke-NUS Graduate Medical School, Singapore 169857, Singapore. Electronic address: cmrhkm@nccs.com.sg.
authors: Kam M
authors: KM
authors: Hui
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D006528:Carcinoma, Hepatocellular / Q000097:blood / Q000175:diagnosis / Q000235:genetics*; D016000:Cluster Analysis; D015331:Cohort Studies; D004265:DNA Helicases / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics*; D042241:Early Diagnosis; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D008113:Liver Neoplasms / Q000097:blood / Q000175:diagnosis / Q000235:genetics*; D016015:Logistic Models; D008297:Male; D008875:Middle Aged; D015999:Multivariate Analysis; D020411:Oligonucleotide Array Sequence Analysis; D010190:Pancreatic Neoplasms / Q000097:blood / Q000175:diagnosis / Q000235:genetics; D054390:Receptors, CCR2 / Q000235:genetics*; D023063:Receptors, Interleukin-8B / Q000235:genetics*; D012680:Sensitivity and Specificity; D013274:Stomach Neoplasms / Q000097:blood / Q000175:diagnosis / Q000235:genetics; D059467:Transcriptome; D055815:Young Adult
chemical_list: C515334:CCR2 protein, human; D004268:DNA-Binding Proteins; D054390:Receptors, CCR2; D023063:Receptors, Interleukin-8B; D004265:DNA Helicases; C498558:EP400 protein, human
keywords: Alpha-fetoprotein (AFP); Blood-borne gene signature; Chemokine (C-X-C motif) receptor 2 (CXCR2); Chronic hepatitis B (CHB); C–C chemokine receptor type 2 (CCR2); E1A-Binding Protein P400 (EP400); Human hepatocellular carcinoma (HCC); Multivariate logistic regression; Peripheral blood mononuclear cells (PBMC)