- Email: [email protected]
264 Matrilin-2, a novel extracellular matrix component in cirrhosis and hepatocellular carcinoma
Category 3." Liver Tumors (Epidemiology, Diagnosis, Management) Results: Pre-incubation of HepO2 cell with 1 mM butyrate acid for 48 h increased efficiency of CD95 (anti-APO1 antibody treatment for 24h) mediated apoptosis (viability 66 vs. 38%). Incubation with butyrate acid alone did not reduce cell viability (98% vital cells after 72h), however strongly induced p53 as a possible molecular mediator of tile observed sensitizing effect. Concomitant treatment with valproic acid (100ug/ml) showed interestingly also an effect on sensitivity towards CD95 mediated apoptosis (35% viability vs. 65% with antibody alone) as well as towards treatment with epirubicine (74 vs. 90% viability). Caspase-3 activity was significantly enhanced in cells treated with valproic acid plus anti CD95 compared to cells treated with antibody. Summary: HDAC inhibitors are a promising class for treatment o f leukemias and solid tumors. Among other class members, valproic acid deserves further evaluation as treatment option for patients with HCC.
[26~ MATRILIN-2, A NOVEL EXTRACELLULAR MATRIX COMPONENT IN CIRRHOSIS AND HEPATOCELLULAR CARCINOMA E. Szabo I , C.S. Paska 1, E Desk 3, I. Kiss3, E. Batmunld{, C.S. Lodi 1, A. Holczbauer 1, A. Kiss I , I. Kovalszky2, Z.S. S c h a ~ . :IL Department
of Pathology, Semmelweis University, Budapest, Hungary," 2I. Department of Pathology and Experimental Cancer Research, Semmelweis Univers#y. Budapest, Hungary; 3Institute of Biochemistry. Biological Research Center of the Hungarian Academy of Sciences, Szeged, Hungary Background and Aims: The recently described matrilin protein family is a part of tile extracellular matrix, whose exact localization, distribution, production and biologic role are still not clarified. It has been detected in embryonic organs, including liver. However, there are no data about matrilins in normal adult liver and pathologic conditions in the liver. The aim of our study was to analyse tile expression of matrilin-2 in normal adult human liver, in cirrhosis and hepatocellular carcinoma. Methods: Formalin fixed, paraflfin-embedded tissue samples from 7 normal human livers, 12 hepatocellular carcinomas (HCC) were used for immunohistochemistry, by using polyclonal rabbit antibody to detect matrilin-2. Six of the non-tumorous surrounding tissue showed cirrhosis, mRNA expression of matrilin-2 was analysed in 12 HCC and surrounding tissue samples fixed in RNA later by real-time PCR. Relative quantification was performed using f3-actin as internal control. Western blot analysis was also used to show the presence ofmatrilin-2. The co-localization of matrilin-2 and laminin was examined by confocal laser scanning microscopy. Results: hnmunohistochemistry showed matrilin-2 expression in tile norreal liver exclusively around the bile ducts, blood vessels and cenwal veins but not in sinusoids. On the other hand intensive protein expression was seen along the sinusoids in cirrhotic livers. The majority of neovasculature in HCC stained positively too. Matrilin-2 colocalized with laminin in all samples studied. No significant differences were found between normal liver and tumor matrilin-2 mRNA expression. Western Not analysis also revealed tile presence of matrilin-2 in liver, however tile results did not show pronounced alteration in protein quantity between normal, cirrhotic and tumor liver. Conclusion: Matrilin-2 is not expressed in the normal liver sinusoids, in contrast to the strong expression in the "capillarized sinusoids" of the cirrhotic liver and neovaseulamre of the tumors. Matrilin-2 was colocalized with laminin. These data indicate that matrilin-2 might be among tile first basement membrane components, which are synthetised during sinusoidal "capillarization" in the cirrhosis and during tumorigenesis. Further, matrilin-2 might be a potential target to influence vascularization. Tile project was supported by grants: Bio14/2001, NKFP-1/0023/2002, ETT-228/2001, OTKA-T037838.
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DIAGNOSIS OF BENIGN AND MALIGNANT PORTAL VEIN THROMBOSIS BY CONTRAST-ENHANCED US IN PATIENTS WITH HCC ON CIRRHOSIS
L. Tarantino 2, G. Francica3, P. Sperlongano I , D. P a r m ~ a n i I , E Esposito4, I. Sordelli I , 1.2 Parmeg~m]ani~. 1 V Division of Surgery -
H University of Naples, Naples, Italy," 2Division of Internal Medicine S. Giovanni di Dio Hospital, Frattamaggiore, Italy," 3Division of Medicine - S. Maria della Pietd Hospital, Casoria, Italy," 4Radiology Depar#nent- Santobono Hospital, Naples, Italy Purpose: To assess the role of contrast-enhanced US (CEUS) in the differential diagnosis between benign and malignant portal vein thrombosis in cirrhotic patients with Hepatocellular carcinoma (HCC). Method and Materials: 54 consecutive patients with drrhosis and biopsy proven HCC, showing thrombosis of main portal vein and/or left and/or right portal vein at US, were prospectively studied with color Doppler US (CD) and CEUS. CEUS was performed at low mechanical index after IV administration of a second-generation contrast agent (SonoVueTM, Bracco, Milan, Italy). Presence or absence of (a) CD signals or (lo) thrombus enhancement at CEUS were considered diagnostic for malignant or benign portal vein thrombosis. 28/54 patients also underwent percutaneous portal vein fine needle biopsy under US g~tidance (FNB). All patients were followed-up bimonthly by CD. Shrinkage of tile thrombus and/or recanalization of the vessels at CD during follow-up were considered definitive evidence of benign nature of the thrombosis while enlargement of the thrombus, disruption of tile vessel's wall and parenchymal infiltration over follow-up were considered consistent with malignancy. CD, CEUS and FNB results were compared to results at follow-up. Results: Follow-up (4-21 months) showed signs of malignant thrombosis in 34/54 patients. FNB gave a true positive result for malignancy in 19/25 patients, a false negative result in 6/25 and a true negative result in 3/3 patients. CD was positive in 7/54 patients. CEUS showed enhancement of tile fllrombus in 30/54 patients. No false positive result was observed at CD, CEUS and FNB. Sensitivity o f CD, CEUS and FNB in detecting malignant thrombi was 20%, 88% and 76%, respectively. 3 patients showed negative CD and CEUS and positive FNB; follow-up confirmed malignant thrombosis in these 3 patients. One patient showed negative CD, CEUS and FNB. Howewer, follow-up o f the thrombus showed US signs of malignancy. A further FNB confirmed HCC infiltration of the portal vein. Conclusions: CELTS seems to be the most sensitive and specific test for diagnosing malignant portal vein thrombosis in patients with cirrhosis.
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ASSESSMENT OF EFFICACY OF PERCUTANEOUS ABLATION OF HEPATOCELLULAR CARCINOMA REAL-TIME CONTRAST-ENHANCED US
L. Tarantino I , G. Francica3, P. Sperlongano2, D. Parmeggiani 2, L. Caserta 3, A. Schiano 3, I. Sordelli 2, U. Parmeggiani 2 . IHepatology
and Interventional US Unit X Giovanni di Dio Hospital ASL NA3, Frattamaggiore, Italy," 2 V Division of Surgery, Second Unioers#y 0 Naples, Naples, Italy," SDivision of Medicine, S. Maria La Pietd Hospital, Casoria, Italy Purpose: Triphasic contrast-enhanced helical CT (CT) is the standard technique for the assessment of results of Percutaneous Ablation Therapies (PATs). The afin of our study was to compare the diagnostic accuracy of Contrast-Enhanced Ultrasound (CEUS) and CT in the assessment of short-term results of Hepatocellular Carcinoma (HCC) ablation. Method and Materials: Fifty-six consecutive cirrhotic patients with a single biopsy proven HCC's nodule (diameter, 1.8-5.0 cm; mean, 3.2 crn) underwent PATs. CT and CEUS were performed in all patients within 7 days before treatment, and within 14 and 21 days after treatment. CELTS was performed at low mechanical index after IV administration of a second-generation contrast agent (BR1, SonoVueTM, Bracco, Milan,
[26~ MATRILIN-2, A NOVEL EXTRACELLULAR MATRIX COMPONENT IN CIRRHOSIS AND HEPATOCELLULAR CARCINOMA E. Szabo I , C.S. Paska 1, E Desk 3, I. Kiss3, E. Batmunld{, C.S. Lodi 1, A. Holczbauer 1, A. Kiss I , I. Kovalszky2, Z.S. S c h a ~ . :IL Department
of Pathology, Semmelweis University, Budapest, Hungary," 2I. Department of Pathology and Experimental Cancer Research, Semmelweis Univers#y. Budapest, Hungary; 3Institute of Biochemistry. Biological Research Center of the Hungarian Academy of Sciences, Szeged, Hungary Background and Aims: The recently described matrilin protein family is a part of tile extracellular matrix, whose exact localization, distribution, production and biologic role are still not clarified. It has been detected in embryonic organs, including liver. However, there are no data about matrilins in normal adult liver and pathologic conditions in the liver. The aim of our study was to analyse tile expression of matrilin-2 in normal adult human liver, in cirrhosis and hepatocellular carcinoma. Methods: Formalin fixed, paraflfin-embedded tissue samples from 7 normal human livers, 12 hepatocellular carcinomas (HCC) were used for immunohistochemistry, by using polyclonal rabbit antibody to detect matrilin-2. Six of the non-tumorous surrounding tissue showed cirrhosis, mRNA expression of matrilin-2 was analysed in 12 HCC and surrounding tissue samples fixed in RNA later by real-time PCR. Relative quantification was performed using f3-actin as internal control. Western blot analysis was also used to show the presence ofmatrilin-2. The co-localization of matrilin-2 and laminin was examined by confocal laser scanning microscopy. Results: hnmunohistochemistry showed matrilin-2 expression in tile norreal liver exclusively around the bile ducts, blood vessels and cenwal veins but not in sinusoids. On the other hand intensive protein expression was seen along the sinusoids in cirrhotic livers. The majority of neovasculature in HCC stained positively too. Matrilin-2 colocalized with laminin in all samples studied. No significant differences were found between normal liver and tumor matrilin-2 mRNA expression. Western Not analysis also revealed tile presence of matrilin-2 in liver, however tile results did not show pronounced alteration in protein quantity between normal, cirrhotic and tumor liver. Conclusion: Matrilin-2 is not expressed in the normal liver sinusoids, in contrast to the strong expression in the "capillarized sinusoids" of the cirrhotic liver and neovaseulamre of the tumors. Matrilin-2 was colocalized with laminin. These data indicate that matrilin-2 might be among tile first basement membrane components, which are synthetised during sinusoidal "capillarization" in the cirrhosis and during tumorigenesis. Further, matrilin-2 might be a potential target to influence vascularization. Tile project was supported by grants: Bio14/2001, NKFP-1/0023/2002, ETT-228/2001, OTKA-T037838.
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DIAGNOSIS OF BENIGN AND MALIGNANT PORTAL VEIN THROMBOSIS BY CONTRAST-ENHANCED US IN PATIENTS WITH HCC ON CIRRHOSIS
L. Tarantino 2, G. Francica3, P. Sperlongano I , D. P a r m ~ a n i I , E Esposito4, I. Sordelli I , 1.2 Parmeg~m]ani~. 1 V Division of Surgery -
H University of Naples, Naples, Italy," 2Division of Internal Medicine S. Giovanni di Dio Hospital, Frattamaggiore, Italy," 3Division of Medicine - S. Maria della Pietd Hospital, Casoria, Italy," 4Radiology Depar#nent- Santobono Hospital, Naples, Italy Purpose: To assess the role of contrast-enhanced US (CEUS) in the differential diagnosis between benign and malignant portal vein thrombosis in cirrhotic patients with Hepatocellular carcinoma (HCC). Method and Materials: 54 consecutive patients with drrhosis and biopsy proven HCC, showing thrombosis of main portal vein and/or left and/or right portal vein at US, were prospectively studied with color Doppler US (CD) and CEUS. CEUS was performed at low mechanical index after IV administration of a second-generation contrast agent (SonoVueTM, Bracco, Milan, Italy). Presence or absence of (a) CD signals or (lo) thrombus enhancement at CEUS were considered diagnostic for malignant or benign portal vein thrombosis. 28/54 patients also underwent percutaneous portal vein fine needle biopsy under US g~tidance (FNB). All patients were followed-up bimonthly by CD. Shrinkage of tile thrombus and/or recanalization of the vessels at CD during follow-up were considered definitive evidence of benign nature of the thrombosis while enlargement of the thrombus, disruption of tile vessel's wall and parenchymal infiltration over follow-up were considered consistent with malignancy. CD, CEUS and FNB results were compared to results at follow-up. Results: Follow-up (4-21 months) showed signs of malignant thrombosis in 34/54 patients. FNB gave a true positive result for malignancy in 19/25 patients, a false negative result in 6/25 and a true negative result in 3/3 patients. CD was positive in 7/54 patients. CEUS showed enhancement of tile fllrombus in 30/54 patients. No false positive result was observed at CD, CEUS and FNB. Sensitivity o f CD, CEUS and FNB in detecting malignant thrombi was 20%, 88% and 76%, respectively. 3 patients showed negative CD and CEUS and positive FNB; follow-up confirmed malignant thrombosis in these 3 patients. One patient showed negative CD, CEUS and FNB. Howewer, follow-up o f the thrombus showed US signs of malignancy. A further FNB confirmed HCC infiltration of the portal vein. Conclusions: CELTS seems to be the most sensitive and specific test for diagnosing malignant portal vein thrombosis in patients with cirrhosis.
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ASSESSMENT OF EFFICACY OF PERCUTANEOUS ABLATION OF HEPATOCELLULAR CARCINOMA REAL-TIME CONTRAST-ENHANCED US
L. Tarantino I , G. Francica3, P. Sperlongano2, D. Parmeggiani 2, L. Caserta 3, A. Schiano 3, I. Sordelli 2, U. Parmeggiani 2 . IHepatology
and Interventional US Unit X Giovanni di Dio Hospital ASL NA3, Frattamaggiore, Italy," 2 V Division of Surgery, Second Unioers#y 0 Naples, Naples, Italy," SDivision of Medicine, S. Maria La Pietd Hospital, Casoria, Italy Purpose: Triphasic contrast-enhanced helical CT (CT) is the standard technique for the assessment of results of Percutaneous Ablation Therapies (PATs). The afin of our study was to compare the diagnostic accuracy of Contrast-Enhanced Ultrasound (CEUS) and CT in the assessment of short-term results of Hepatocellular Carcinoma (HCC) ablation. Method and Materials: Fifty-six consecutive cirrhotic patients with a single biopsy proven HCC's nodule (diameter, 1.8-5.0 cm; mean, 3.2 crn) underwent PATs. CT and CEUS were performed in all patients within 7 days before treatment, and within 14 and 21 days after treatment. CELTS was performed at low mechanical index after IV administration of a second-generation contrast agent (BR1, SonoVueTM, Bracco, Milan,