A molecular signature to discriminate dysplastic nodules from early hepatocellular carcinoma in HCV cirrhosis.
OBJECTIVE
Small liver nodules approximately 2 cm are difficult to characterize by radiologic or pathologic examination. Our aim was to identify a molecular signature to diagnose early hepatocellular carcinoma (HCC).


METHODS
The transcriptional profiles of 55 candidate genes were assessed by quantitative real-time reverse-transcription polymerase chain reaction (RT-PCR) in 17 dysplastic nodules (diameter, 10 mm) and 20 early HCC (diameter, 18 mm) from HCV cirrhotic patients undergoing resection/transplantation and 10 nontumoral cirrhotic tissues and 10 normal liver tissues. Candidate genes were confirmed by quantitative RT-PCR in 20 advanced HCCs and by immunohistochemistry in 75 samples and validated in an independent set of 29 samples (dysplastic nodules [10] and small HCC [19; diameter, 20 mm]).


RESULTS
Twelve genes were significantly, differentially expressed in early HCCs compared with dysplastic nodules (>2-fold change; area under the receiver operating characteristic curve > or =0.8): this included TERT, GPC3, gankyrin, survivin, TOP2A, LYVE1, E-cadherin, IGFBP3, PDGFRA, TGFA, cyclin D1, and HGF. Logistic regression analysis identified a 3-gene set including GPC3 (18-fold increase in HCC, P = .01), LYVE1 (12-fold decrease in HCC, P = .0001), and survivin (2.2-fold increase in HCC, P = .02), which had a discriminative accuracy of 94%. The validity of the gene signature was confirmed in a prospective testing set. GPC3 immunostaining was positive in all HCCs and negative in dysplastic nodules (22/22 vs 0/14, respectively, P < .001). Nuclear staining for survivin was positive in 12 of 13 advanced HCC cases and in 1 of 9 early tumors.


CONCLUSIONS
Molecular data based on gene transcriptional profiles of a 3-gene set allow a reliable diagnosis of early HCC. Immunostaining of GPC3 confirms the diagnosis of HCC.
Llovet
Josep M
JM
Mount Sinai Liver Cancer Program, Department of Medicine, Mount Sinai School of Medicine, New York 10029, USA. Josep.Llovet@mssm.edu
Chen
Yingbei
Y
Wurmbach
Elisa
E
Roayaie
Sasan
S
Fiel
M Isabel
MI
Schwartz
Myron
M
Thung
Swan N
SN
Khitrov
Gregory
G
Zhang
Weijia
W
Villanueva
Augusto
A
Battiston
Carlo
C
Mazzaferro
Vincenzo
V
Bruix
Jordi
J
Waxman
Samuel
S
Friedman
Scott L
SL
D014408:Biomarkers, Tumor / Q000235:genetics; D006528:Carcinoma, Hepatocellular / Q000175:diagnosis* / Q000209:etiology / Q000235:genetics* / Q000473:pathology; D004273:DNA, Neoplasm / Q000235:genetics; D003937:Diagnosis, Differential; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D052138:Genes, Neoplasm / Q000235:genetics; D006023:Glycoproteins / Q000235:genetics; D053673:Glypicans / Q000235:genetics; D016174:Hepacivirus; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins; D008103:Liver Cirrhosis / Q000150:complications / Q000175:diagnosis* / Q000473:pathology / Q000821:virology*; D008107:Liver Diseases / Q000175:diagnosis / Q000235:genetics; D008113:Liver Neoplasms / Q000175:diagnosis* / Q000209:etiology / Q000235:genetics* / Q000473:pathology; D008869:Microtubule-Associated Proteins / Q000235:genetics; D009363:Neoplasm Proteins / Q000235:genetics; D000077022:Survivin; D033921:Vesicular Transport Proteins
C107489:BIRC5 protein, human; D014408:Biomarkers, Tumor; D004273:DNA, Neoplasm; C105937:GPC3 protein, human; D006023:Glycoproteins; D053673:Glypicans; D051034:Inhibitor of Apoptosis Proteins; C119248:LYVE1 protein, human; D008869:Microtubule-Associated Proteins; D009363:Neoplasm Proteins; D000077022:Survivin; D033921:Vesicular Transport Proteins