- Email: [email protected]
118 ADVANCED CYTOLOGY FOR THE DIAGNOSIS OF MALIGNANT BILE DUCT STRICTURES IN PATIENTS WITH JAUNDICE: A TWO-STEP APPROACH
ORAL PRESENTATIONS (p = 0.05) score: ≤3 = 0, or ≥4 = 2. Two prognostic groups were identified in the TC according to score values ≤2 or ≥3, with 5-yr risk of recurrence of 16.7±2.6% and 47.5±4.7%, respectively (p < 0.0001). The model was validated in the 2 validation cohorts with, in particular, a 5-yr risk of recurrence of 9.0±1.7% (score ≤2) vs 47.3±9.5% (score ≥3), p < 0.001, in the ABM VC. In both cohorts, a score ≤2 was associated with better 5-yr overall survival: RVC: 80.6±3.5% vs 66.7±10.3% (p = 0.06) and ABM VC: 67.7±3.4% vs 49.1±7.4% (p = 0.002). In patients with score ≤2, recurrence and survival were similar whether patients met or not Milan criteria. Conclusions: 1. AFP level is highly predictive of HCC recurrence after LT. 2. A prognostic score including AFP significantly improves the identification of patients with expanded criteria at low risk of recurrence. 3. A score ≤2 could be proposed for selection of these patients. 116 USING DYNAMIC CONTRAST-ENHANCED MAGNETIC RESONANCE IMAGING (DCE-MRI) TO PREDICT TREATMENT OUTCOMES FOR ADVANCED HEPATOCELLULAR CARCINOMA (HCC) PATIENTS WHO RECEIVED SORAFENIB PLUS TEGAFUR/URACIL THERAPY Y.C. Shen1,2,3 , C.Y. Hsu4 , C.W. Yu4 , C. Hsu1,2 , C.H. Hsu1,2 , A.L. Cheng1,2,5 , T.F. Shih4 . 1 Department of Oncology, 2 National Center of Excellence for Clinical Trial and Research, 3 Department of Medical Research, 4 Department of Medical Imaging, 5 Department of Internal Medicine, National Taiwan University Hospital, Taipei, Taiwan R.O.C. E-mail: [email protected] Background and Aims: Anti-angiogenesis is considered an important mechanism for the anti-tumor effects of sorafenib and metronomic chemotherapy in HCC. This study evaluated the correlation of vascular response measured by DCE-MRI and the clinical outcomes of advanced HCC patients who were treated with sorafenib plus metronomic tegafur/uracil. Methods: Advanced HCC patients who were enrolled in a phase II trial of sorafenib (800 mg/day) and tegafur/uracil (250 mg/m2 /day, based on tegafur) and agreed with additional DCE-MRI studies were eligible. DCE-MRI was performed before and after 3, 14, and 56 days of study treatment. An operator-defined region of interest was placed in the most strongly enhanced area of the tumor in all patients to obtain the pharmacokinetic parameter Ktrans , which reflected the tumor vascular permeability and perfusion. The “vascular response”, defined as a decline in Ktrans after treatment by ≥40%, was correlated with the best tumor response (by the response evaluation criteria in solid tumors, RECIST) and progression-free survival (PFS). Correlation of vascular response and outcomes Vascular response (Ktrans decrease ≥40%) Yes Tumor response by RECIST (no. of patients) PR/SD 12 PD 1 p-value1 Progression-free survival (weeks) Median (95% CI) 29.1 (10.6, 47.6) p-value2 1
No 7 11 0.003 8.7 (7.7, 9.7) 0.0237
Fisher’s exact test; 2 Log-rank test.
Results: Forty-one patients were enrolled and data from 31 evaluable patients (27 men, 4 women; median age 55.9 years) were analyzed. The clinical characteristics of the analyzed patients were not different from that of all patients. One (3.2%), 18 (58.1%) and 12 (38.7%) patients had partial response (PR), stable disease S52
(SD) and progressive disease (PD), respectively. The Ktrans change after 14-day study treatment was found the most informative: Ktrans decreased by 44.7% ±29.0% in patients with PR/SD and increased by 16.2% ± 35.6% in those with PD (p-value <0.0005). Correlation of vascular response with RECIST response and PFS is summarized in the table. Conclusion: The vascular response after 14-day treatment correlates well with tumor response and PFS in HCC patients who received sorafenib/tegafur/uracil therapy. Ktrans changes measured by DCE-MRI after 14-day treatment may be used as a predictive biomarker for bioactivity and efficacy of anti-angiogenic therapy. Supported by grants NSC97–3112-B-002–012, NSC98–3112-B-002– 007 and NCTRC200711. 117 SEROMARKERS FOR LONG-TERM PREDICTION OF HEPATOCELLULAR CARCINOMA RISK IN PATIENTS WITH CHRONIC HEPATITIS C M.-H. Lee1,2 , H.-I. Yang2 , S.-N. Lu3 , C.-L. Jen2 , S.-H. Yeh1 , C.-J. Liu1 , P.-J. Chen1 , S.-L. You2 , L.-Y. Wang4 , W.-J. Chen1 , C.-J. Chen1,2 , R.E.V.E.A.L-HCV Study Group. 1 National Taiwan University, 2 Genomics Research Center, Academia Sinica, Taipei, 3 Kaohsiung Chang-Gung Memorial Hospital, Kaohsiung, 4 MacKay Medical College, Taipei, Taiwan R.O.C. E-mail: [email protected] Background: Hepatitis C virus (HCV) contributes to one-third of hepatocellular carcinoma cases worldwide. Long-term predictors for HCV-related hepatocellular carcinoma are essential for early intervention. Serum HCV RNA levels, alanine aminotransferase (ALT) levels and HCV genotype were assessed for their predictability of hepatocellular carcinoma risk. Methods: A prospective cohort of 936 participants positive for antibodies against HCV and aged 30–65 years were recruited from seven townships in Taiwan and followed from 1991 to 2006. Serum HCV RNA level, ALT, and HCV genotypes were examined at enrollment or during follow-up. Newly-developed hepatocellular carcinoma was identified by health examination and computerized linkage with national cancer registration and death certification profiles. Multivariate-adjusted hazard ratios with 95% confidence intervals were estimated by Cox regression models. Results: There were 59 newly developed hepatocellular carcinoma cases during 8,561 person-years of follow-up, giving an incidence rate of 689.2 per 100,000 person-years. The cumulative risks for hepatocellular carcinoma were 1.1% for HCV RNA seronegative, 8.7% for low HCV RNA levels, and 14.4% for high HCV RNA levels, in correspondingly (p < 0.001). Patients had increased cumulative risk for hepatocellular carcinoma with elevated serum ALT, with 1.7% for persistently ≤15 U/L, 4.4% for ever >15 U/L but never >45 U/L, and 14.4% for ALT ever >45 U/L (p < 0.001). HCV genotype 1 had higher risk than HCV genotype non-1, with cumulative risk of 13.0% and 4.5% (p < 0.001). Conclusions: Patients with elevated serum HCV RNA levels, ALT levels, or HCV genotype 1 are independent risk predictors of hepatocellular carcinoma. These results have strong implications for the treatment of chronic hepatitis C infection. 118 ADVANCED CYTOLOGY FOR THE DIAGNOSIS OF MALIGNANT BILE DUCT STRICTURES IN PATIENTS WITH JAUNDICE: A TWO-STEP APPROACH M. Sartori1 , M. Orsello1 , F. Montino1 , M. Ballare´ 1 , M. Pagliarulo1 , R. Boldorini2 , A. Paganotti2 , S. Carmagnola1 , S. Andorno1 , M. Del Piano1 . 1 AOU Maggiore della Carit` a, 2 University of Eastern Piedmont ‘A. Avogadro’, Novara, Italy E-mail: [email protected] Background and Aims: Fluorescence in situ hybridization (FISH), an ancillary cytologic technique, increases the sensitivity for detecting
Journal of Hepatology 2010 vol. 52 | S43–S54
ORAL PRESENTATIONS pancreatobiliary tract cancer over routine cytology. The aim of this study was to assess the diagnostic value of FISH for the detection of malignancy in biliary tract strictures of patients with extrahepatic jaundice in whom routine cytology tested negative for cancer. Patients and Methods: Brush cytological specimens from 65 consecutive jaundiced patients (37 males and 28 females, mean age 69±12 years) who underwent endoscopic retrograde cholangiopancreatography for pancreatobiliary strictures were examined by routine cytology and FISH. With FISH, specimens were considered positive for malignancy if at least 5 polysomic cells, or at least ten trisomy of chromosome 7 or 3 were present. Definitive diagnosis of the stricture as benign or malignant was based on surgical pathology (30 cases) or sufficient clinical and radiological follow up >12 months (35 cases). Results: Forty-six of 65 patients had surgical pathologic and/or clinical-radiological evidence of malignancy (23 cholangiocarcinomas, 20 pancreatic carcinomas, 3 other malignant tumors) while 19 had benign strictures. Routine cytology showed 56% sensitivity but 100% specificity, 100% PPV and 47% NPV for the diagnosis of malignancy, while FISH showed 87% sensitivity, 95% specificity, 97% PPV and 75% NPV. In 39 patients with routine cytologic specimens negative for malignancy (20 of whom had indeed cancer), the sensitivity, specificity, PPV and NPV for malignancy of FISH were respectively 75%, 94%, 94%, and 77%. Conclusions: FISH is recommended as a second step to detect cancer in jaundiced patients with pancreatobiliary tract strictures and routine cytologic specimens negative for malignancy. 119 IDENTIFICATION OF RESECTED BENIGN HEPATOCELLULAR TUMORS MAY REQUIRE MORE THAN ROUTINE PATHOLOGICAL MARKERS P. Bioulac-Sage1,2 , H. Laumonier3 , C. Laurent4 , G. Cubel5 , A. Sa Cunha6 , J. Saric4 , C. Balabaud1,2,7 , J. Zucman-Rossi8 . 1 Pathology, CHU Bordeaux, 2 Inserm U889, Universit´e Bordeaux2, 3 Radiology, 4 Surgery, CHU Bordeaux, 5 Inserm U889, Universit´e Bordeaux2, Bordeaux, 6 Surgery, CHU Bordeaux, Pessac, 7 Hepatology, CHU Bordeaux, Bordeaux, 8 Inserm U674, Universit´e Paris Descartes, Paris, France E-mail: [email protected] Background and Aims: Identification of focal nodular hyperplasia (FNH) and hepatocellular adenoma (HCA) is usually easy on the resected specimen using routine techniques (H&E, trichrome, reticulin and eventually CK7, CK19, CD34, glypican 3). However, as a tertiary referral centre, we have to interpret difficult cases with problems of differential diagnosis between FNH, HCA, and well differentiated hepatocellular carcinoma (wd-HCC), particularly nodules with steatosis or sinusoidal dilatation. The aim of our study was to evaluate the usefulness of the new set of markers1 in daily practice. Methods: Markers were used retrospectively (from 2000 for FNH and 1990 for HCA) and prospectively from 2007 up to now (92 FNH, 141 HCA). Immunostainings included glutamine synthetase (GS), liver fatty acid binding protein (LFABP), serum amyloid A (SAA), C-reactiv protein (CRP) and b-catenin (b-cat). GS staining defined FNH when a characteristic map-like pattern was present and b-cat activated HCA (b-cat HCA) when diffusely/strongly positive, associated with aberrant nuclear b-cat positivity; LFABP negative was characteristic of HNF1a-inactivated HCA (H-HCA) and SAA/CRP + of inflammatory (I)HCA. Results: Using GS, the diagnosis of FNH was straightforward in all cases but one. Using the complete set of markers, the diagnosis of HCA was straightforward in 90% of cases. LFABP was sometimes difficult to interpret when the staining in the non tumoral liver (NTL) was faint; SAA/CRP when there was a staining in NTL (after bleeding, portal vein embolization). HCA cases with diffuse/weak or heterogeneous positivity of GS and no or a very few nuclear
b-cat staining may require molecular biology for b-cat mutations confirmation. In some cases, it was difficult to differentiate these b-cat HCA from wd HCC except if the last ones express glypican 3. Markers modified the diagnosis in 7 cases: from FNH to HCA (5 cases), from HCA to FNH (2 cases). In addition markers allowed the classification of HCA in H-HCA, IHCA and b-cat HCA. Conclusion: These results suggest that these markers should be used in tertiary centres to identify with greater certainty and easiness FNH and HCA, and subsequently HCA subtypes. Reference(s) [1] Bioulac-Sage. Hepatology 2007.
120 A 7 GENE SET ASSOCIATED WITH CHRONIC HYPOXIA OF UNIVERSAL PROGNOSTIC IMPORTANCE IN HEPATOCELLULAR CARCINOMA H. van Malenstein1 , O. Gevaert2 , L. Libbrecht1 , A. Daemen2 , J. Allemeersch3 , F. Nevens1 , E. Van Cutsem4 , D. Cassiman1 , B. De Moor2 , C. Verslype1 , J. van Pelt1 . 1 Hepatology, University Hospital Gasthuisberg, 2 ESAT, 3 VIB, Catholic University, 4 Digestive Oncology, University Hospital Gasthuisberg, Leuven, Belgium E-mail: [email protected] Background and Aims: Hepatocellular carcinomas (HCCs) are heterogeneous tumors with an unpredictable clinical course. There is a need for more objective prognostic criteria to decide on treatment options. Molecular classification of HCCs might provide better insight in prognosis and patient directed therapy. Since the behavior of solid tumors depends in part on tumor microenvironment, we hypothesized that in HCC certain regions exist with a characteristic gene expression related to chronic hypoxia which will induce aggressive behavior. Methods: We determined the gene expression pattern for human HepG2 liver cells under chronic hypoxia by microarray (20% O2 vs. 2% O2 during 72 hrs). Significant differentially expressed genes were selected and their clinical value was assessed. In our hypothesis-driven analysis we included available independent microarray studies of patients with HCC in one single analysis. Three microarray studies were used as training sets to determine a minimal prognostic gene set associated with poor prognostic indicators and one additional study was used for validation. Results: Using computational methods we identified 7 genes, out of 3592 differentially expressed under chronic hypoxia, that showed correlation with poor prognosis in all training sets (272 patients) and this was validated in a 4th dataset (91 patients). The expression of the 7 gene set was liver specific compared to other tissues, as tested in published data and in other cell lines. In a separate analysis on one of the four training sets, the 7-gene set is associated with poor survival (HR 1.39, p = 0.007) and early recurrence (HR 2.92, p = 0.007). Retrospectively, using a hypoxia score based on this 7-gene set we found that patients with a score >0.35 had a median survival of 307 days, whereas patients with a score ≤0.35 had a median survival of 1602 days (p = 0.005). Conclusions: In our analysis we could include available microarray studies of patients with HCC in one single analysis, irrespective of etiology, diagnostic parameters or molecular technique used. We identified a unique, liver specific 7-gene signature associated with chronic hypoxia that correlates with poor prognosis in HCCs.
Journal of Hepatology 2010 vol. 52 | S43–S54
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No 7 11 0.003 8.7 (7.7, 9.7) 0.0237
Fisher’s exact test; 2 Log-rank test.
Results: Forty-one patients were enrolled and data from 31 evaluable patients (27 men, 4 women; median age 55.9 years) were analyzed. The clinical characteristics of the analyzed patients were not different from that of all patients. One (3.2%), 18 (58.1%) and 12 (38.7%) patients had partial response (PR), stable disease S52
(SD) and progressive disease (PD), respectively. The Ktrans change after 14-day study treatment was found the most informative: Ktrans decreased by 44.7% ±29.0% in patients with PR/SD and increased by 16.2% ± 35.6% in those with PD (p-value <0.0005). Correlation of vascular response with RECIST response and PFS is summarized in the table. Conclusion: The vascular response after 14-day treatment correlates well with tumor response and PFS in HCC patients who received sorafenib/tegafur/uracil therapy. Ktrans changes measured by DCE-MRI after 14-day treatment may be used as a predictive biomarker for bioactivity and efficacy of anti-angiogenic therapy. Supported by grants NSC97–3112-B-002–012, NSC98–3112-B-002– 007 and NCTRC200711. 117 SEROMARKERS FOR LONG-TERM PREDICTION OF HEPATOCELLULAR CARCINOMA RISK IN PATIENTS WITH CHRONIC HEPATITIS C M.-H. Lee1,2 , H.-I. Yang2 , S.-N. Lu3 , C.-L. Jen2 , S.-H. Yeh1 , C.-J. Liu1 , P.-J. Chen1 , S.-L. You2 , L.-Y. Wang4 , W.-J. Chen1 , C.-J. Chen1,2 , R.E.V.E.A.L-HCV Study Group. 1 National Taiwan University, 2 Genomics Research Center, Academia Sinica, Taipei, 3 Kaohsiung Chang-Gung Memorial Hospital, Kaohsiung, 4 MacKay Medical College, Taipei, Taiwan R.O.C. E-mail: [email protected] Background: Hepatitis C virus (HCV) contributes to one-third of hepatocellular carcinoma cases worldwide. Long-term predictors for HCV-related hepatocellular carcinoma are essential for early intervention. Serum HCV RNA levels, alanine aminotransferase (ALT) levels and HCV genotype were assessed for their predictability of hepatocellular carcinoma risk. Methods: A prospective cohort of 936 participants positive for antibodies against HCV and aged 30–65 years were recruited from seven townships in Taiwan and followed from 1991 to 2006. Serum HCV RNA level, ALT, and HCV genotypes were examined at enrollment or during follow-up. Newly-developed hepatocellular carcinoma was identified by health examination and computerized linkage with national cancer registration and death certification profiles. Multivariate-adjusted hazard ratios with 95% confidence intervals were estimated by Cox regression models. Results: There were 59 newly developed hepatocellular carcinoma cases during 8,561 person-years of follow-up, giving an incidence rate of 689.2 per 100,000 person-years. The cumulative risks for hepatocellular carcinoma were 1.1% for HCV RNA seronegative, 8.7% for low HCV RNA levels, and 14.4% for high HCV RNA levels, in correspondingly (p < 0.001). Patients had increased cumulative risk for hepatocellular carcinoma with elevated serum ALT, with 1.7% for persistently ≤15 U/L, 4.4% for ever >15 U/L but never >45 U/L, and 14.4% for ALT ever >45 U/L (p < 0.001). HCV genotype 1 had higher risk than HCV genotype non-1, with cumulative risk of 13.0% and 4.5% (p < 0.001). Conclusions: Patients with elevated serum HCV RNA levels, ALT levels, or HCV genotype 1 are independent risk predictors of hepatocellular carcinoma. These results have strong implications for the treatment of chronic hepatitis C infection. 118 ADVANCED CYTOLOGY FOR THE DIAGNOSIS OF MALIGNANT BILE DUCT STRICTURES IN PATIENTS WITH JAUNDICE: A TWO-STEP APPROACH M. Sartori1 , M. Orsello1 , F. Montino1 , M. Ballare´ 1 , M. Pagliarulo1 , R. Boldorini2 , A. Paganotti2 , S. Carmagnola1 , S. Andorno1 , M. Del Piano1 . 1 AOU Maggiore della Carit` a, 2 University of Eastern Piedmont ‘A. Avogadro’, Novara, Italy E-mail: [email protected] Background and Aims: Fluorescence in situ hybridization (FISH), an ancillary cytologic technique, increases the sensitivity for detecting
Journal of Hepatology 2010 vol. 52 | S43–S54
ORAL PRESENTATIONS pancreatobiliary tract cancer over routine cytology. The aim of this study was to assess the diagnostic value of FISH for the detection of malignancy in biliary tract strictures of patients with extrahepatic jaundice in whom routine cytology tested negative for cancer. Patients and Methods: Brush cytological specimens from 65 consecutive jaundiced patients (37 males and 28 females, mean age 69±12 years) who underwent endoscopic retrograde cholangiopancreatography for pancreatobiliary strictures were examined by routine cytology and FISH. With FISH, specimens were considered positive for malignancy if at least 5 polysomic cells, or at least ten trisomy of chromosome 7 or 3 were present. Definitive diagnosis of the stricture as benign or malignant was based on surgical pathology (30 cases) or sufficient clinical and radiological follow up >12 months (35 cases). Results: Forty-six of 65 patients had surgical pathologic and/or clinical-radiological evidence of malignancy (23 cholangiocarcinomas, 20 pancreatic carcinomas, 3 other malignant tumors) while 19 had benign strictures. Routine cytology showed 56% sensitivity but 100% specificity, 100% PPV and 47% NPV for the diagnosis of malignancy, while FISH showed 87% sensitivity, 95% specificity, 97% PPV and 75% NPV. In 39 patients with routine cytologic specimens negative for malignancy (20 of whom had indeed cancer), the sensitivity, specificity, PPV and NPV for malignancy of FISH were respectively 75%, 94%, 94%, and 77%. Conclusions: FISH is recommended as a second step to detect cancer in jaundiced patients with pancreatobiliary tract strictures and routine cytologic specimens negative for malignancy. 119 IDENTIFICATION OF RESECTED BENIGN HEPATOCELLULAR TUMORS MAY REQUIRE MORE THAN ROUTINE PATHOLOGICAL MARKERS P. Bioulac-Sage1,2 , H. Laumonier3 , C. Laurent4 , G. Cubel5 , A. Sa Cunha6 , J. Saric4 , C. Balabaud1,2,7 , J. Zucman-Rossi8 . 1 Pathology, CHU Bordeaux, 2 Inserm U889, Universit´e Bordeaux2, 3 Radiology, 4 Surgery, CHU Bordeaux, 5 Inserm U889, Universit´e Bordeaux2, Bordeaux, 6 Surgery, CHU Bordeaux, Pessac, 7 Hepatology, CHU Bordeaux, Bordeaux, 8 Inserm U674, Universit´e Paris Descartes, Paris, France E-mail: [email protected] Background and Aims: Identification of focal nodular hyperplasia (FNH) and hepatocellular adenoma (HCA) is usually easy on the resected specimen using routine techniques (H&E, trichrome, reticulin and eventually CK7, CK19, CD34, glypican 3). However, as a tertiary referral centre, we have to interpret difficult cases with problems of differential diagnosis between FNH, HCA, and well differentiated hepatocellular carcinoma (wd-HCC), particularly nodules with steatosis or sinusoidal dilatation. The aim of our study was to evaluate the usefulness of the new set of markers1 in daily practice. Methods: Markers were used retrospectively (from 2000 for FNH and 1990 for HCA) and prospectively from 2007 up to now (92 FNH, 141 HCA). Immunostainings included glutamine synthetase (GS), liver fatty acid binding protein (LFABP), serum amyloid A (SAA), C-reactiv protein (CRP) and b-catenin (b-cat). GS staining defined FNH when a characteristic map-like pattern was present and b-cat activated HCA (b-cat HCA) when diffusely/strongly positive, associated with aberrant nuclear b-cat positivity; LFABP negative was characteristic of HNF1a-inactivated HCA (H-HCA) and SAA/CRP + of inflammatory (I)HCA. Results: Using GS, the diagnosis of FNH was straightforward in all cases but one. Using the complete set of markers, the diagnosis of HCA was straightforward in 90% of cases. LFABP was sometimes difficult to interpret when the staining in the non tumoral liver (NTL) was faint; SAA/CRP when there was a staining in NTL (after bleeding, portal vein embolization). HCA cases with diffuse/weak or heterogeneous positivity of GS and no or a very few nuclear
b-cat staining may require molecular biology for b-cat mutations confirmation. In some cases, it was difficult to differentiate these b-cat HCA from wd HCC except if the last ones express glypican 3. Markers modified the diagnosis in 7 cases: from FNH to HCA (5 cases), from HCA to FNH (2 cases). In addition markers allowed the classification of HCA in H-HCA, IHCA and b-cat HCA. Conclusion: These results suggest that these markers should be used in tertiary centres to identify with greater certainty and easiness FNH and HCA, and subsequently HCA subtypes. Reference(s) [1] Bioulac-Sage. Hepatology 2007.
120 A 7 GENE SET ASSOCIATED WITH CHRONIC HYPOXIA OF UNIVERSAL PROGNOSTIC IMPORTANCE IN HEPATOCELLULAR CARCINOMA H. van Malenstein1 , O. Gevaert2 , L. Libbrecht1 , A. Daemen2 , J. Allemeersch3 , F. Nevens1 , E. Van Cutsem4 , D. Cassiman1 , B. De Moor2 , C. Verslype1 , J. van Pelt1 . 1 Hepatology, University Hospital Gasthuisberg, 2 ESAT, 3 VIB, Catholic University, 4 Digestive Oncology, University Hospital Gasthuisberg, Leuven, Belgium E-mail: [email protected] Background and Aims: Hepatocellular carcinomas (HCCs) are heterogeneous tumors with an unpredictable clinical course. There is a need for more objective prognostic criteria to decide on treatment options. Molecular classification of HCCs might provide better insight in prognosis and patient directed therapy. Since the behavior of solid tumors depends in part on tumor microenvironment, we hypothesized that in HCC certain regions exist with a characteristic gene expression related to chronic hypoxia which will induce aggressive behavior. Methods: We determined the gene expression pattern for human HepG2 liver cells under chronic hypoxia by microarray (20% O2 vs. 2% O2 during 72 hrs). Significant differentially expressed genes were selected and their clinical value was assessed. In our hypothesis-driven analysis we included available independent microarray studies of patients with HCC in one single analysis. Three microarray studies were used as training sets to determine a minimal prognostic gene set associated with poor prognostic indicators and one additional study was used for validation. Results: Using computational methods we identified 7 genes, out of 3592 differentially expressed under chronic hypoxia, that showed correlation with poor prognosis in all training sets (272 patients) and this was validated in a 4th dataset (91 patients). The expression of the 7 gene set was liver specific compared to other tissues, as tested in published data and in other cell lines. In a separate analysis on one of the four training sets, the 7-gene set is associated with poor survival (HR 1.39, p = 0.007) and early recurrence (HR 2.92, p = 0.007). Retrospectively, using a hypoxia score based on this 7-gene set we found that patients with a score >0.35 had a median survival of 307 days, whereas patients with a score ≤0.35 had a median survival of 1602 days (p = 0.005). Conclusions: In our analysis we could include available microarray studies of patients with HCC in one single analysis, irrespective of etiology, diagnostic parameters or molecular technique used. We identified a unique, liver specific 7-gene signature associated with chronic hypoxia that correlates with poor prognosis in HCCs.
Journal of Hepatology 2010 vol. 52 | S43–S54
S53