- Email: [email protected]
Hepatocellular cancer gene expression profiling by DNA microarrays: diagnostic and therapeutic implications
Poster Sessions
94
Results: 411/623 (66%) answered. 96% routinely screened HCC with ultrasonography (98%) and alphafetoprotein assay (95%) at 6 (77%), 3 (14%) or 12-month intervals (13%). Screening was applied to cirrhosis (100%) or extensive fibrosis (54%), independently of etiology (80%) and Child score (61%), but according to age (49%) and curative treatment feasibility (43%). In case of detection of a small hypoechogenic nodule in a young patient with compensated HCV-cirrhosis, 234 performed histological examination. Proposed treatments were resection (49%), transplantation (30%) or percutaneous alcohol injection (PEI) (16%) in case of biopsy-proven HCC; a second biopsy (47%), treatment (either PEI, resection or transplantation) (21%) or ultrasonographic follow-up (21%) were proposed in case of non biopsy-proven HCC. 90% wished a prospective screening study comparing etheir no screening versus screening (430/o),or 2 periodicities of screening (55%) limited to Child A or B cirrhosis patients t70 years. Conclusions: Most of French specialists routinely screen cirrhotic patients for HCC, but with rather different modalities. In case of small HCC without contraindication to curative treatment, surgical resection is performed in half of patients.
I
153
INVOLVEMENT OF MATRIX METALLOPROTEINASES IN HEPATOCYTE GROWTH FACTOR-MDUCED INVASION OF HUMAN HEPATOCELLULAR CARCINOMA CELLS IN VITRO
Amaud Monvoisin, Jean Rosenbaum. Vniversite Victor Segalen Bordeaux, France
Intra-hepatic invasion is a key feature of hepatocellular carcinoma (HCC) progression. We have shown that human liver myofibroblasts, a cell type infiltrating HCC stroma, induce in vitro invasion of HCC cells via the secretion of hepatccyte growth factor (HGF). This effect could be reproduced by recombinant HGF. As matrix invasion, requires proteolytic degradation of its components, we investigated the role of matrix metalloprotenaises (MMP) family members in HGF-induced HCC cells invasion. We first examined MMP expression by the HCC cell line HepG2, using RT-PCR. MMPs 1, 7, 8, 9, and 10 were not expressed in HepG2 cells. MMP-2 transcripts were detected but were not regulated by HGF. MMP-3 transcripts were undetectable in unstimulated HepG2 cells but became clearly expressed in HGF-stimulated cells. Northern blot analysis confirmed the induction of MMP3 mRNA expression in HGF-stimulated HepG2 cells. By Western blot, we found that HGF dose-dependently stimulated the secretion of pro-MMP-3 in the culture medium. Involvement of MMPs in HGF-induced invasion was confirmed with the use of inhibitors. Marimastat, a synthetic MMP inhibitor decreased HGF-induced invasion of HepG2 cells with a maximum of 82.7 f 13.3% at 20 PM. TIMP-2, a natural inhibitor, also decreased invasion up to 51.2 f 11.2% at 200 ng/ml. Finally, RT-PCR demonstrated MMP3 expression in most human HCCs tested. In conclusion, our data demonstrate that MMPs, most likely MMP-3, mediate HGF-induced invasion of HCC cells. The in vivo expression of MMP-3 in HCC suggests a role for this protease in HCC progression.
I
154
A POSIBLE LINK BETWEEN POLYAMINES AND THIOL REDOX SIGNALING PATWAYS IN DIABETIC LIVER
D.D. Pavlovic, G. Bjelakovic. Institute of Biochemistry Faculty of Medicine, Nis, Yugoslavia
It was shown that the activation of PKC as well as MAPK by hyperglycemia, AGE products and ROS may affect redox-senzitive signaling pathway of the cell, that can be. responsible for liver failure in diabetes including alteration of cellular phenotype, growth and apoptosis. On the other hand intracellular thiols are crucial in the control of NF-KB activity + a classic redox sensor, and low thiol levels are required for induction of this transcription factor. Since we have previously demonstrated that polyamines possess free radicals scavenging properties, the aim of this
study was to see whether any linkage exists between redox status and polyamine content in liver of diabetic animals. Methods: Oxidative stress was evaluated by the quantification of MDA and reduced glutathione (GSH) content. Fractions of polyamines were determined by electrophoresis. The rats were allocated to the following groups: I-control; II-treated with single dose of streptozotocine St (50 mg/kg bw); III-treated simultaneously with spermidine (50 mmol/kg bw) ip. 1 h before administration of St as well as continually with single daily dose following 15 days); IV-treated with the Spd only. The results in liver tissue and serum are summarized in the tables. Liver tissue Variable
Control streptozotocine St + spd Spd GSH (FmoVg pmt.) 3.57f0.86 1.26&0.62*** 3.78f0.6 4.3f0.73** MDA (nmol/mg prot.) 11.3f0.7 18.24f2.5”’ 12.7f1.7”“” 11.7f1.23 Putrescine (put) nmol/g 190f32 140&24* 2OOf45” 26Of18” Spermine (Sp) nmol/g 770f118 420f89*** 820f124”“” 810f116 Suermidine (SDd)nmoYe 1020f286 660+94*** 1030f226”“” 134of218’ Serum
Glucose (nmolil) 6.55 f 0.9 30.2 f 7.14*** 9.36 f 2.7***“” 6.42 f 0.97 AST (u/l) 121.4 f 20 335.3 f 58*** 164.5 f 15’“” 139.8 f 15.3 *p < 0.05; ** < 0.01; ***p < 0.001 vs. control; “p < 0.01; On < 0.001 vs. streptozotocine treated rats The decrease of GSH concentration in liver of diabetic animals may lead to alteration in the redox state which is principally buffered by GSH. It was also shown that polyamines inhibit PKC and that depletion of polyamines might increase activity of cJun amino-terminal kinases (JNKs) (type of MAPK) as well as that polyamines might be involved in the caspase activating apoptosis signal cascade. Based on these findings as well as on the fact that the concentrations of GSH and polyamines were reversed upon simultaneous treatment by St and Spd, and that Spd given alone significantly increased the content of GSH, we postulated that there is possible link between polyamines and down regulation of redox cell signalization. Taken together our results suggest that spermidine might prevent diabetic liver damage through a thiol redox mechanism that has practical as well as theoretical implications. 208 HEPATOCELLULAR CANCER GENE EXPRESSION L-J PROFILING BY DNA MICROARRAYS: DIAGNOSTIC AND THERAPEUTIC IMPLICATIONS S. Kaiser, H. Hass, M. Gregor. Department of Medicine, Universiv of Tuebingen, Germany
Both chronic hepatitis B and C share the risk of development of hepatocellular carcinoma (HCC) as the severest complication. To elucidate differential changes in gene expression in the pathogenesis of HCC, tissue transcripts from hepatitis B and C induced HCCs were profiled using high-density oligonucleotide microarrays. RNA from tumor and non-malignant liver tissue was used to prepare cRNA for hybridization to oligonucleotide DNA chips. GeneChips were analyzed and changes > 3 fold over control were evaluated. Hepatitis B and C induced HCC tissues displayed remarkably different gene expression profiles, which were assigned to 8 categories. 96 genes in hepatitis B induced HCC and 37 in hepatitis C induced HCC were overexpressed, while 73 in hepatitis B and 69 in hepatitis C were downregulated. Both novel and known ‘HCC-related’ genes, including those for cell cycle regulators, apoptosis-related genes, detoxification genes, defensins, metalloproteinases, cytochrome p450 family and collagens were differentially regulated. Unlike hepatitis B induced HCC, hepatitis C induced HCC appears as an epithelial cancer sharing many common changes in global gene expression profile with other tumors such as colon cancer with major changes in inflammatory-mediator, cytokine/growth factor and cancerrelated categories. In contrast, hepatitis B induced HCC displays many unique genetic features not observed in other tumor profiles. Microarray
Category 4: Hepatocellular
carcinoma,
expression profiling demonstrates, for the first time, broad and fundamental differences in pathogenic mechanisms of hepatitis B versus C induced HCC. These studies will aid in developing specific tumor markers and targeted therapies for HCC and identify candidate genes for genome-wide susceptibility gene searches.
I
287
HEMANGIOMA-LIKE LESIONS IN PATIENTS WITH CHRONIC LIVER DISEASE: DIAGNOSTIC EVALUATION
E. Caturelli, M. Pompili. D.A. Siena, A. Andriulli, M. Bisceglia. U.O. Gastroenterologia, Ospetie “Casa Sollievo della Sofferenza” WCCS, Cattedra di Medicina Interna II, Universitd Cattolica de1 Sacro Cuore, Roma; V.O. Gastroenterologia, Ospedale “Casa Sollievo della Sofferenza” IRCCS, San Giovanni Rotondo (Fg); U.O. Gastroenterologia, Ospedale “Casa Sollievo della Sofferenza” IRCCS, San Giovanni Rotondo (Fg); Servizio Anatomia e Istologia Patologica gia, Ospedale “Casa Sollievo della Sofferenza” IRCCS, San Giovanni Rotondo (Fg), Italy Purpose:
To quantify the risk of misdiagnosis of focal hepatic lesions presenting on ultrasonography (US) as “typical hemangiomas” in a population at high risk for HCC and to identify the most effective approach to their differential diagnosis. Materials and Methods 1982 newly diagnosed cirrhotics underwent US and serum alpha fetoprotein (AFP) studies for early HCC detection. Focal lesions with typical features of hemangioma were subjected to confirmatory contrast-enhanced dynamic or spiral computed tomography (CT) and/or technetium 99m-labeled red blood cell single photon emission CT and (in the absence of image confirmation) US-guided fine-needle biopsy. Patients whose initial scan revealed no lesions or lesions diagnosed as hemangiomas were enrolled in an US follow-up program. All hemangioma-like lesions emerged during follow-up were studied as described above or (during the last 3 years of the study) biopsied directly. Results: Initial US revealed hemangioma-like lesions in 44 out of 1982 patients: 22 were hemangiomas, and 22 were HCC. The 26 hemangioma-like lesions detected during me follow-up of 1648 patients were 22 HCC and 4 dysplastic nodules. No patient with hemangioma-like lesion presented elevated AFP serum levels. Conclusions: If the initial US examination of a cirrhotic liver reveals a “typical hemangioma”, confirmatory imaging studies are necessary since 50% (in our study) will be hyperechoic HCCs. The probability of HCC (or preneoplastic lesions) is virtually 100% for hemangioma-like lesions detected on US along the follow-up program. US-guided biopsy can be safely performed to confirm the diagnosis. Diagnostic AFP serum levels are not found in patients with hyperechoic HCC.
I 288
DIAGNOSIS OF LIVER NODULES OBSERVED IN CIRRHOTICS DURING ULTRASOUND SCREENING FOR EARLY DETECTION OF HEPATOCELLULAR CARCINOMA
E. Caturelli, D.A. Siena, M. Vigliotti, V. Attino, M. Bisceglia, A. Andriulli. U.O. Gastroenterologia.. U.O. Ematologia., Servizio Anatomia E Istologia Patologica., Servizio Anatomia E Istologia Patologica., U.O. Gastroenterologia, Italy Purpose: To evaluate the nature of focal liver lesions detected during the ultrasound follow-up of a cirrhotic (prevalently anti-HCV positive) population. Materials and Methods: The study population consisted in 1827 consecutive newly diagnosed cirrhotic patients without liver nodules at enrollment. Patients were screened at 4-month intervals by ultrasound (US) and serum alpha fetoprotein (AFP) assessment. All lesions detected on imaging studies (except those accompanied by significantly elevated AFP levels and those with hyperechoic appearance) were subjected to biopsy (histology and cytology).
liver regeneration,
apoptosis
95
Results: During the 7-year follow-up period (mean 43.1 months), one or more solid focal lesions were found in 287 patients. AFP was diagnostic for hepatocellular carcinoma (HCC) in 51 patients. US-guided fine-needle biopsy was performed in the remaining 236 patients, yielding a diagnosis in 214: 198 HCC, 11 adenomatous hyperplasias, and five B cell non-Hodgkin lymphomas (all confined to the liver and all in patients with HCV-related disease). Twenty-two patients with non-diagnostic biopsies received diagnoses of HCC (20) or adenomatous hyperplasia (2) based on arteriography or surgical biopsy. Conclusions: Focal lesions arising in patients with HCV-related liver cirrhosis can be other than HCC, and US-guided fine-needle biopsy plays an important role in their diagnosis. The prevalence of non-Hodgkin lymphoma in this selected population was 0.31%. The fact that all five lymphoma patients had cirrhosis related to hepatitis C strengthens the hypothesis of an etiologic correlation between the latter infection and B cell lymphoproliferative disorders.
I 289
APOPTOSIS VERSUS CYTOPROLIFERATION IN HEPATOCELLULAR CARCINOMA: ROLE OF PROTEIN OXIDATION
E.H. Ibrahim, H.A. El-Aggan, S.A. Mahmoud, L.K. Younis. Epatobiliary Unit, Department of Medicine, Faculty of Medicine, University of Alexandria, Egypt
Objective: Neoplastic growth is determined by the balance between cell death and cell proliferation, the present study was designed to study the rates of apoptosis and proliferation in hepatocellular carcinoma (HCC) in relation to tumor growth and protein oxidation. Methods: Fifty patients with cirrhosis (30 with histologically-proven HCC and 20 without HCC) were included in the study. In HCC patients, core liver biopsies were analyzed for apoptosis using the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nickend labeling method and for ~53 expression by immunohistochemistry. The proliferative activity of HCC was assessed using proliferating cell nuclear antigen labeling. In all patients, the plasma levels of carbonyl proteins and advanced oxidation protein products, markers of protein oxidation, and plasma xanthine oxidase activity were determined. Results: In HCC, both the apoptotic rate and the proliferative activity increased with decreased tumor differentiation and with increasing tumor size and stage @ < 0.01). A positive correlation was found between the proliferative activity and both the apoptotic rate and ~53 expression @ < 0.01). However, the apoptotic rate was always less than the proliferative activity and their ratio tended to decrease in higher tumor grade, size and stage and in p53positive tumors @ 5 0.001). Meanwhile, The plasma oxidized proteins levels and the plasma xanthine oxidase activity showed a significant stepwise increase in cirrhotic patients without HCC and with the subsequent development of HCC (p < 0.0001). The markers of protein oxidation in HCC patients had positive correlations with the proliferative activity and inverse correlations with the ratio between the apoptotic rate and the proliferative activity @ < 0.05), but showed no significant correlation with the apoptotic rate @ < 0.05). Conclusions: Cell proliferation predominates over apoptosis during HCC progression coincident with expression of mutant ~53 and an enhanced protein oxidation. Also, oxidative stress seems to play a role in the progress of liver cirrhosis to HCC
I 294
ANGIOGENIN IN HEPATOCELLULAR CARCINOMA: CORRELATION WlTH TUMOR VASCULARITY AND PROLIFERATIVE ACTIVITY
H.A. El Aggan, S.E. Hegab, N.M. Baddour, A.F. Ketat. Hepatobiliary Unit, Department of Medicine, Faculty of Medicine, University of Alexandria, Egypt
Objective: Angiogenesis is an essential process for growth and metas-
94
Results: 411/623 (66%) answered. 96% routinely screened HCC with ultrasonography (98%) and alphafetoprotein assay (95%) at 6 (77%), 3 (14%) or 12-month intervals (13%). Screening was applied to cirrhosis (100%) or extensive fibrosis (54%), independently of etiology (80%) and Child score (61%), but according to age (49%) and curative treatment feasibility (43%). In case of detection of a small hypoechogenic nodule in a young patient with compensated HCV-cirrhosis, 234 performed histological examination. Proposed treatments were resection (49%), transplantation (30%) or percutaneous alcohol injection (PEI) (16%) in case of biopsy-proven HCC; a second biopsy (47%), treatment (either PEI, resection or transplantation) (21%) or ultrasonographic follow-up (21%) were proposed in case of non biopsy-proven HCC. 90% wished a prospective screening study comparing etheir no screening versus screening (430/o),or 2 periodicities of screening (55%) limited to Child A or B cirrhosis patients t70 years. Conclusions: Most of French specialists routinely screen cirrhotic patients for HCC, but with rather different modalities. In case of small HCC without contraindication to curative treatment, surgical resection is performed in half of patients.
I
153
INVOLVEMENT OF MATRIX METALLOPROTEINASES IN HEPATOCYTE GROWTH FACTOR-MDUCED INVASION OF HUMAN HEPATOCELLULAR CARCINOMA CELLS IN VITRO
Amaud Monvoisin, Jean Rosenbaum. Vniversite Victor Segalen Bordeaux, France
Intra-hepatic invasion is a key feature of hepatocellular carcinoma (HCC) progression. We have shown that human liver myofibroblasts, a cell type infiltrating HCC stroma, induce in vitro invasion of HCC cells via the secretion of hepatccyte growth factor (HGF). This effect could be reproduced by recombinant HGF. As matrix invasion, requires proteolytic degradation of its components, we investigated the role of matrix metalloprotenaises (MMP) family members in HGF-induced HCC cells invasion. We first examined MMP expression by the HCC cell line HepG2, using RT-PCR. MMPs 1, 7, 8, 9, and 10 were not expressed in HepG2 cells. MMP-2 transcripts were detected but were not regulated by HGF. MMP-3 transcripts were undetectable in unstimulated HepG2 cells but became clearly expressed in HGF-stimulated cells. Northern blot analysis confirmed the induction of MMP3 mRNA expression in HGF-stimulated HepG2 cells. By Western blot, we found that HGF dose-dependently stimulated the secretion of pro-MMP-3 in the culture medium. Involvement of MMPs in HGF-induced invasion was confirmed with the use of inhibitors. Marimastat, a synthetic MMP inhibitor decreased HGF-induced invasion of HepG2 cells with a maximum of 82.7 f 13.3% at 20 PM. TIMP-2, a natural inhibitor, also decreased invasion up to 51.2 f 11.2% at 200 ng/ml. Finally, RT-PCR demonstrated MMP3 expression in most human HCCs tested. In conclusion, our data demonstrate that MMPs, most likely MMP-3, mediate HGF-induced invasion of HCC cells. The in vivo expression of MMP-3 in HCC suggests a role for this protease in HCC progression.
I
154
A POSIBLE LINK BETWEEN POLYAMINES AND THIOL REDOX SIGNALING PATWAYS IN DIABETIC LIVER
D.D. Pavlovic, G. Bjelakovic. Institute of Biochemistry Faculty of Medicine, Nis, Yugoslavia
It was shown that the activation of PKC as well as MAPK by hyperglycemia, AGE products and ROS may affect redox-senzitive signaling pathway of the cell, that can be. responsible for liver failure in diabetes including alteration of cellular phenotype, growth and apoptosis. On the other hand intracellular thiols are crucial in the control of NF-KB activity + a classic redox sensor, and low thiol levels are required for induction of this transcription factor. Since we have previously demonstrated that polyamines possess free radicals scavenging properties, the aim of this
study was to see whether any linkage exists between redox status and polyamine content in liver of diabetic animals. Methods: Oxidative stress was evaluated by the quantification of MDA and reduced glutathione (GSH) content. Fractions of polyamines were determined by electrophoresis. The rats were allocated to the following groups: I-control; II-treated with single dose of streptozotocine St (50 mg/kg bw); III-treated simultaneously with spermidine (50 mmol/kg bw) ip. 1 h before administration of St as well as continually with single daily dose following 15 days); IV-treated with the Spd only. The results in liver tissue and serum are summarized in the tables. Liver tissue Variable
Control streptozotocine St + spd Spd GSH (FmoVg pmt.) 3.57f0.86 1.26&0.62*** 3.78f0.6 4.3f0.73** MDA (nmol/mg prot.) 11.3f0.7 18.24f2.5”’ 12.7f1.7”“” 11.7f1.23 Putrescine (put) nmol/g 190f32 140&24* 2OOf45” 26Of18” Spermine (Sp) nmol/g 770f118 420f89*** 820f124”“” 810f116 Suermidine (SDd)nmoYe 1020f286 660+94*** 1030f226”“” 134of218’ Serum
Glucose (nmolil) 6.55 f 0.9 30.2 f 7.14*** 9.36 f 2.7***“” 6.42 f 0.97 AST (u/l) 121.4 f 20 335.3 f 58*** 164.5 f 15’“” 139.8 f 15.3 *p < 0.05; ** < 0.01; ***p < 0.001 vs. control; “p < 0.01; On < 0.001 vs. streptozotocine treated rats The decrease of GSH concentration in liver of diabetic animals may lead to alteration in the redox state which is principally buffered by GSH. It was also shown that polyamines inhibit PKC and that depletion of polyamines might increase activity of cJun amino-terminal kinases (JNKs) (type of MAPK) as well as that polyamines might be involved in the caspase activating apoptosis signal cascade. Based on these findings as well as on the fact that the concentrations of GSH and polyamines were reversed upon simultaneous treatment by St and Spd, and that Spd given alone significantly increased the content of GSH, we postulated that there is possible link between polyamines and down regulation of redox cell signalization. Taken together our results suggest that spermidine might prevent diabetic liver damage through a thiol redox mechanism that has practical as well as theoretical implications. 208 HEPATOCELLULAR CANCER GENE EXPRESSION L-J PROFILING BY DNA MICROARRAYS: DIAGNOSTIC AND THERAPEUTIC IMPLICATIONS S. Kaiser, H. Hass, M. Gregor. Department of Medicine, Universiv of Tuebingen, Germany
Both chronic hepatitis B and C share the risk of development of hepatocellular carcinoma (HCC) as the severest complication. To elucidate differential changes in gene expression in the pathogenesis of HCC, tissue transcripts from hepatitis B and C induced HCCs were profiled using high-density oligonucleotide microarrays. RNA from tumor and non-malignant liver tissue was used to prepare cRNA for hybridization to oligonucleotide DNA chips. GeneChips were analyzed and changes > 3 fold over control were evaluated. Hepatitis B and C induced HCC tissues displayed remarkably different gene expression profiles, which were assigned to 8 categories. 96 genes in hepatitis B induced HCC and 37 in hepatitis C induced HCC were overexpressed, while 73 in hepatitis B and 69 in hepatitis C were downregulated. Both novel and known ‘HCC-related’ genes, including those for cell cycle regulators, apoptosis-related genes, detoxification genes, defensins, metalloproteinases, cytochrome p450 family and collagens were differentially regulated. Unlike hepatitis B induced HCC, hepatitis C induced HCC appears as an epithelial cancer sharing many common changes in global gene expression profile with other tumors such as colon cancer with major changes in inflammatory-mediator, cytokine/growth factor and cancerrelated categories. In contrast, hepatitis B induced HCC displays many unique genetic features not observed in other tumor profiles. Microarray
Category 4: Hepatocellular
carcinoma,
expression profiling demonstrates, for the first time, broad and fundamental differences in pathogenic mechanisms of hepatitis B versus C induced HCC. These studies will aid in developing specific tumor markers and targeted therapies for HCC and identify candidate genes for genome-wide susceptibility gene searches.
I
287
HEMANGIOMA-LIKE LESIONS IN PATIENTS WITH CHRONIC LIVER DISEASE: DIAGNOSTIC EVALUATION
E. Caturelli, M. Pompili. D.A. Siena, A. Andriulli, M. Bisceglia. U.O. Gastroenterologia, Ospetie “Casa Sollievo della Sofferenza” WCCS, Cattedra di Medicina Interna II, Universitd Cattolica de1 Sacro Cuore, Roma; V.O. Gastroenterologia, Ospedale “Casa Sollievo della Sofferenza” IRCCS, San Giovanni Rotondo (Fg); U.O. Gastroenterologia, Ospedale “Casa Sollievo della Sofferenza” IRCCS, San Giovanni Rotondo (Fg); Servizio Anatomia e Istologia Patologica gia, Ospedale “Casa Sollievo della Sofferenza” IRCCS, San Giovanni Rotondo (Fg), Italy Purpose:
To quantify the risk of misdiagnosis of focal hepatic lesions presenting on ultrasonography (US) as “typical hemangiomas” in a population at high risk for HCC and to identify the most effective approach to their differential diagnosis. Materials and Methods 1982 newly diagnosed cirrhotics underwent US and serum alpha fetoprotein (AFP) studies for early HCC detection. Focal lesions with typical features of hemangioma were subjected to confirmatory contrast-enhanced dynamic or spiral computed tomography (CT) and/or technetium 99m-labeled red blood cell single photon emission CT and (in the absence of image confirmation) US-guided fine-needle biopsy. Patients whose initial scan revealed no lesions or lesions diagnosed as hemangiomas were enrolled in an US follow-up program. All hemangioma-like lesions emerged during follow-up were studied as described above or (during the last 3 years of the study) biopsied directly. Results: Initial US revealed hemangioma-like lesions in 44 out of 1982 patients: 22 were hemangiomas, and 22 were HCC. The 26 hemangioma-like lesions detected during me follow-up of 1648 patients were 22 HCC and 4 dysplastic nodules. No patient with hemangioma-like lesion presented elevated AFP serum levels. Conclusions: If the initial US examination of a cirrhotic liver reveals a “typical hemangioma”, confirmatory imaging studies are necessary since 50% (in our study) will be hyperechoic HCCs. The probability of HCC (or preneoplastic lesions) is virtually 100% for hemangioma-like lesions detected on US along the follow-up program. US-guided biopsy can be safely performed to confirm the diagnosis. Diagnostic AFP serum levels are not found in patients with hyperechoic HCC.
I 288
DIAGNOSIS OF LIVER NODULES OBSERVED IN CIRRHOTICS DURING ULTRASOUND SCREENING FOR EARLY DETECTION OF HEPATOCELLULAR CARCINOMA
E. Caturelli, D.A. Siena, M. Vigliotti, V. Attino, M. Bisceglia, A. Andriulli. U.O. Gastroenterologia.. U.O. Ematologia., Servizio Anatomia E Istologia Patologica., Servizio Anatomia E Istologia Patologica., U.O. Gastroenterologia, Italy Purpose: To evaluate the nature of focal liver lesions detected during the ultrasound follow-up of a cirrhotic (prevalently anti-HCV positive) population. Materials and Methods: The study population consisted in 1827 consecutive newly diagnosed cirrhotic patients without liver nodules at enrollment. Patients were screened at 4-month intervals by ultrasound (US) and serum alpha fetoprotein (AFP) assessment. All lesions detected on imaging studies (except those accompanied by significantly elevated AFP levels and those with hyperechoic appearance) were subjected to biopsy (histology and cytology).
liver regeneration,
apoptosis
95
Results: During the 7-year follow-up period (mean 43.1 months), one or more solid focal lesions were found in 287 patients. AFP was diagnostic for hepatocellular carcinoma (HCC) in 51 patients. US-guided fine-needle biopsy was performed in the remaining 236 patients, yielding a diagnosis in 214: 198 HCC, 11 adenomatous hyperplasias, and five B cell non-Hodgkin lymphomas (all confined to the liver and all in patients with HCV-related disease). Twenty-two patients with non-diagnostic biopsies received diagnoses of HCC (20) or adenomatous hyperplasia (2) based on arteriography or surgical biopsy. Conclusions: Focal lesions arising in patients with HCV-related liver cirrhosis can be other than HCC, and US-guided fine-needle biopsy plays an important role in their diagnosis. The prevalence of non-Hodgkin lymphoma in this selected population was 0.31%. The fact that all five lymphoma patients had cirrhosis related to hepatitis C strengthens the hypothesis of an etiologic correlation between the latter infection and B cell lymphoproliferative disorders.
I 289
APOPTOSIS VERSUS CYTOPROLIFERATION IN HEPATOCELLULAR CARCINOMA: ROLE OF PROTEIN OXIDATION
E.H. Ibrahim, H.A. El-Aggan, S.A. Mahmoud, L.K. Younis. Epatobiliary Unit, Department of Medicine, Faculty of Medicine, University of Alexandria, Egypt
Objective: Neoplastic growth is determined by the balance between cell death and cell proliferation, the present study was designed to study the rates of apoptosis and proliferation in hepatocellular carcinoma (HCC) in relation to tumor growth and protein oxidation. Methods: Fifty patients with cirrhosis (30 with histologically-proven HCC and 20 without HCC) were included in the study. In HCC patients, core liver biopsies were analyzed for apoptosis using the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nickend labeling method and for ~53 expression by immunohistochemistry. The proliferative activity of HCC was assessed using proliferating cell nuclear antigen labeling. In all patients, the plasma levels of carbonyl proteins and advanced oxidation protein products, markers of protein oxidation, and plasma xanthine oxidase activity were determined. Results: In HCC, both the apoptotic rate and the proliferative activity increased with decreased tumor differentiation and with increasing tumor size and stage @ < 0.01). A positive correlation was found between the proliferative activity and both the apoptotic rate and ~53 expression @ < 0.01). However, the apoptotic rate was always less than the proliferative activity and their ratio tended to decrease in higher tumor grade, size and stage and in p53positive tumors @ 5 0.001). Meanwhile, The plasma oxidized proteins levels and the plasma xanthine oxidase activity showed a significant stepwise increase in cirrhotic patients without HCC and with the subsequent development of HCC (p < 0.0001). The markers of protein oxidation in HCC patients had positive correlations with the proliferative activity and inverse correlations with the ratio between the apoptotic rate and the proliferative activity @ < 0.05), but showed no significant correlation with the apoptotic rate @ < 0.05). Conclusions: Cell proliferation predominates over apoptosis during HCC progression coincident with expression of mutant ~53 and an enhanced protein oxidation. Also, oxidative stress seems to play a role in the progress of liver cirrhosis to HCC
I 294
ANGIOGENIN IN HEPATOCELLULAR CARCINOMA: CORRELATION WlTH TUMOR VASCULARITY AND PROLIFERATIVE ACTIVITY
H.A. El Aggan, S.E. Hegab, N.M. Baddour, A.F. Ketat. Hepatobiliary Unit, Department of Medicine, Faculty of Medicine, University of Alexandria, Egypt
Objective: Angiogenesis is an essential process for growth and metas-