- Email: [email protected]
566 FLUORESCENCE IN SITU HYBRIDIZATION (FISH) ANALYSIS FOR THE DIAGNOSIS OF HCC
POSTERS Conclusions: AFP has a characteristic to stimulate the expression of CXCR4 in hepatoma cells, this was first evidences to show that AFP up-regulated the expression of CXCR4 through inhibing the activity of PTEN. Acknowledgements: The National Natural Science Foundation of China(No. 30660117, 30760090; 30960153) and The Natural Science Foundation of Hainan Province(No. 309034). 566 FLUORESCENCE IN SITU HYBRIDIZATION (FISH) ANALYSIS FOR THE DIAGNOSIS OF HCC J. Lorber, H. Kreipe, B. Schlegelberger, M.P. Manns, B. Skawran, T.F. Greten. Hannover Medical School, Hannover, Germany E-mail: [email protected] Introduction: Hepatocellular carcinoma (HCC) is the third most common reason for cancer-related death worldwide. According to EASL guidelines HCC can be diagnosed by imaging studies and pathological evaluation of tissue biopsies. However, in some cases pathological examination is very difficult and further studies are need for the exact diagnosis of HCC. In the past, we have described that an increase in chromosomal instability observed by FISH analysis during dedifferentiation of HCC. The aim of this study was to evaluate of FISH analysis can be used as a routine procedure for the diagnosis of HCC in combination with conventional pathological examination. Methods: Tissue biopsies from a total of 63 patients with new intra-hepatic lesions were biopsied and by conventional pathology and fluorescence in situ hybridization (FISH). FISH was performed, using a set of probes specific for the centromeres of chromosomes 1 and 8. Results from both pathological analysis and FISH analysis were compared with clinical outcome of patients in order to verify the correctness of diagnosis. Results: In this retrospective analysis only patients were included, who presented at our institution and for whom, a pathological report, a FISH analysis and the clinical course for a minimum of 18 months was available. In 42/63 cases clinical outcome correlated with both pathology report and FISH analysis. In 2 cases FISH analysis as well as pathology report were negative despite of a positive clinical outcome. In 3/63 cases FISH analysis but not the pathology report were suggestive for HCC, while the clinical outcome was positive and in 14/63 cases pathology report was positive but FISH analysis was negative while the clinical outcome was suggestive for HCC. Finally in 1/63 cases either pathology report or FISH report were suggestive for HCC, while the clinical outcome was negative. Conclusion: This retrospective analysis suggests that FISH analysis can help analyzing biopsies from patients with intra-hepatic lesions suspicious for HCC. However negative results cannot rule out the presence of HCC. 567 USEFULNESS OF MICROFLUIDIC CARDS TO PREDICT RESISTANCE TO ANTICANCER CHEMOTHERAPY IN LIVER TUMOURS E. Lozano1 , P. Martinez-Becerra1 , M.A. Serrano1 , M.R. Romero1 , N. Grane ˜ 2 , L. Trigueros-Motos2 , A. Del Rio3 , I. Monte1 , F.J. Casado2 , 5 M. Molina-Arcas2 , L. Alvarez4 , L. Munoz-Bellvis ˜ , M. PastorAnglada2 , J.J.G. Marin1 . 1 Laboratory of Experimental Hepatology and Drug Targeting, University of Salamanca, CIBERehd, Salamanca, 2 Departament de Bioqu´ımica i Biologia Molecular, Institut de Biomedicina de la Universitat de Barcelona, CIBERehd, Barcelona, 3 Servicio de Aparato Digestivo, Sanatorio Sagrado Coraz´ on de Jesus, Valladolid, 4 Research Unit, La Paz University Hospital, Madrid, 5 Division of Hepatobiliary and Pancreatic Surgery, University Hospital, Salamanca, Spain E-mail: [email protected] Background and Aims: Poor response to chemotherapy is a serious and common problem affecting different types of liver cancer. In
addition to novel anticancer agents, it is also important to develop new tools to predict drug-specific refractoriness of an individual tumour in order to orientate the pharmacological regime with the highest probability of success in the shortest time. Although genetic fingerprint using DNA/RNA microchips provide a large amount of information its interpretation is complex. MicroFluidic Cards (MFCs) constitute a cheaper, faster and easier to interpreting alternative. Methods: 130 genes with potential role in resistance/sensitivity to anticancer drugs were selected from previously published studies. They were classified according to their presumed involvement in the following Mechanisms of Chemoresistance (MOC): Drug uptake (MOC-Ia) or efflux (MOC-Ib); Metabolic pro-drugs activation or active agents inactivation (MOC-II); Change in the expression or activity of molecular targets (MOC-III); DNA-repairing (MOC-IV); and survival/apoptosis balance (MOC-V). Using real-time RT-QPCR with SYBR Green detection, the expression of these genes in healthy tissue and liver tumours was investigated. The result of this firststep analysis has permitted to select 92 (plus three housekeeping) genes to be included in a MFC to determine mRNA abundance by real-time RT-QPCR using Taqman® probes. This has been used in a retrospective pilot study on paired samples of healthy and tumour tissue collected from patients with cholangiocarcinoma, hepatoblastoma or hepatocellular carcinoma (n≥5 each), as well as from liver tumour cells (wild type and chemoresistant sublines) cultured without or with IC50 of usual anticancer drugs. Results: Based on the results a predictive model was developed. The mRNA abundance of each gene and its known role in MOCs was combined to calculate gene resistance coefficient (GRC). Integration of GCRs within each MOC and considering the importance of each MOC in the refractoriness to a given drug or drug family has permits to determine tumour resistance index (TRI). This is a value of predictive resistance of a specific tumour to a drug or family of drugs. Conclusions: A novel tool to predict resistance of liver tumours to chemotherapy has been developed. Preliminary data recommend further validation in prospective studies. 568 A NOVEL PROGNOSTIC ESTIMATION FORMULA FOR HEPATOCELLULAR CARCINOMA AFTER RESECTION, BASED ON TNM STAGING, TOTAL BILIRUBIN LEVEL, BLOOD LOSS, HLA-DR ALPHA AND DNA METHYLTRANSFERASE 1 W. Lu1 , J. Dong1 , P. Bie2 , D. Guo3 , W. Zhang1 , S. Cai1 . 1 The Department of Hepatobiliary Surgery, Chinese PLA General Hospital, Beijing, 2 The Pathology Institute of Southwest Hospital, 3 The Pathology Institute Southwest Hospital, Third Military Medical University, Chongqing, China E-mail: [email protected] Background and Aims: Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide. However, the present prognostic systems (e.g. pathological TNM, CLIP etc.) cannot provide a precise prognosis for individuals. We introduce a novel estimation formula to predict prognosis in HCC patients after resection based on independent prognostic biomarkers and clinical factors. Methods: 234 patients who undergo curative hepatectomy for HCC were retrospect analyzed. Three candidate moleculars (DNA methyltransferase 1, HLA-DR alpha, b-catenin) were tested by tissue microarray (TMA) based immunohistochemistry. The Cox proportional-hazards regression model was used for both univariate and multivariate analyses of all the preoperative, intraoperative, postoperative clinical factors and the three candidate molecules. Then an estimation system to predict prognosis was developed, based on significant independent prognostic biomarkers and clinical factors. The area under the ROC curve was employed to compare the accuracy of the formula, TNM, CLIP, CUPI, Okuda staging systems to predict outcome of HCC after liver resection.
Journal of Hepatology 2010 vol. 52 | S183–S317
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addition to novel anticancer agents, it is also important to develop new tools to predict drug-specific refractoriness of an individual tumour in order to orientate the pharmacological regime with the highest probability of success in the shortest time. Although genetic fingerprint using DNA/RNA microchips provide a large amount of information its interpretation is complex. MicroFluidic Cards (MFCs) constitute a cheaper, faster and easier to interpreting alternative. Methods: 130 genes with potential role in resistance/sensitivity to anticancer drugs were selected from previously published studies. They were classified according to their presumed involvement in the following Mechanisms of Chemoresistance (MOC): Drug uptake (MOC-Ia) or efflux (MOC-Ib); Metabolic pro-drugs activation or active agents inactivation (MOC-II); Change in the expression or activity of molecular targets (MOC-III); DNA-repairing (MOC-IV); and survival/apoptosis balance (MOC-V). Using real-time RT-QPCR with SYBR Green detection, the expression of these genes in healthy tissue and liver tumours was investigated. The result of this firststep analysis has permitted to select 92 (plus three housekeeping) genes to be included in a MFC to determine mRNA abundance by real-time RT-QPCR using Taqman® probes. This has been used in a retrospective pilot study on paired samples of healthy and tumour tissue collected from patients with cholangiocarcinoma, hepatoblastoma or hepatocellular carcinoma (n≥5 each), as well as from liver tumour cells (wild type and chemoresistant sublines) cultured without or with IC50 of usual anticancer drugs. Results: Based on the results a predictive model was developed. The mRNA abundance of each gene and its known role in MOCs was combined to calculate gene resistance coefficient (GRC). Integration of GCRs within each MOC and considering the importance of each MOC in the refractoriness to a given drug or drug family has permits to determine tumour resistance index (TRI). This is a value of predictive resistance of a specific tumour to a drug or family of drugs. Conclusions: A novel tool to predict resistance of liver tumours to chemotherapy has been developed. Preliminary data recommend further validation in prospective studies. 568 A NOVEL PROGNOSTIC ESTIMATION FORMULA FOR HEPATOCELLULAR CARCINOMA AFTER RESECTION, BASED ON TNM STAGING, TOTAL BILIRUBIN LEVEL, BLOOD LOSS, HLA-DR ALPHA AND DNA METHYLTRANSFERASE 1 W. Lu1 , J. Dong1 , P. Bie2 , D. Guo3 , W. Zhang1 , S. Cai1 . 1 The Department of Hepatobiliary Surgery, Chinese PLA General Hospital, Beijing, 2 The Pathology Institute of Southwest Hospital, 3 The Pathology Institute Southwest Hospital, Third Military Medical University, Chongqing, China E-mail: [email protected] Background and Aims: Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide. However, the present prognostic systems (e.g. pathological TNM, CLIP etc.) cannot provide a precise prognosis for individuals. We introduce a novel estimation formula to predict prognosis in HCC patients after resection based on independent prognostic biomarkers and clinical factors. Methods: 234 patients who undergo curative hepatectomy for HCC were retrospect analyzed. Three candidate moleculars (DNA methyltransferase 1, HLA-DR alpha, b-catenin) were tested by tissue microarray (TMA) based immunohistochemistry. The Cox proportional-hazards regression model was used for both univariate and multivariate analyses of all the preoperative, intraoperative, postoperative clinical factors and the three candidate molecules. Then an estimation system to predict prognosis was developed, based on significant independent prognostic biomarkers and clinical factors. The area under the ROC curve was employed to compare the accuracy of the formula, TNM, CLIP, CUPI, Okuda staging systems to predict outcome of HCC after liver resection.
Journal of Hepatology 2010 vol. 52 | S183–S317
S225