- Email: [email protected]
249 Risk factors and patterns of extrahepatic metastasis of hepatocellular carcinoma
03. Liver tumors' (epidemiology, diagnosis', managemen 0
I•
EFFECT OF LOW DOSE 5-FLUOROURACIL AND CISPLATIN INTRA-ARTERIAL INFUSION C H E M O T H E R A P Y IN ADVANCED HEPATOCELLULAR C A R C I N O M A WITH DECOMPENSATED CIRRHOSIS
J.Y. Cheong 1, S.W. Cho 1, T.Y. Lim 1, K.M. Lee 1, K.B. Hahm2, J.H. Kim 1.
1Dep. Gastroenterology, 2Genomic Research Center for Gastroenterology, Ajou University School of Medicine, Suwon, South Korea Background and Aims: Advanced hepatocellular carcinoma (HCC) with portal vein thrombosis (PVT) has a poor prognosis. The aim of this study was to evaluate the efficacy and safety of repeated arterial infusion of low dose cisplatin and 5-fluorouracil (FU) in patients who were diagnosed as advanced HCC with decompensated cirrhosis. Methods: Between January 1995 and December 2003, a total of 79 decompensated cirrhotic patients with HCC having PVT were analyzed and divided into 2 groups. Group 1 (n 40) received intra-arterial infusion chemotherapy with cisplatin (10mg for 5 days) and 5-FU (250mg for 5 days) via implanted chemoport, and group 2 (n 39) was managed with only conservative treatment. Results: The two groups were well matched with respect to features relating to prognosis including age, sex, Chil~Pugh class. Although diffuse tumor involvement, main portal vein tumor thrombosis and bi-lobar involvement were more frequent in group 1, the median survival period of group 1 was significantly longer than group 2 (5 months vs. 3 months, P 0.0156). Also, the 1-year survival rate of group 1 (7.5%) was higher than that of group 2 (5.1%) (P 0.0156). If we analyze the patients with Child class B, survival benefits of intra-arterial chemotherapy were more significant (P 0.0077). Conclusions: Intra-arterial chemotherapy consisting of low dose 5-FU and cisplatin achieved favorable result in advanced HCC patients who had decompensated cirrhosis and showed better survival in selected patients. This therapy may be useful as a palliative treatment for HCC patients in decompensated cirrhosis.
I-~
EFFICACY OF HEPATITIS B VIRUS VACCINATION ON THE REDUCTION OF HEPATOCELLULAR C A R C I N O M A INCIDENCE: POST-NATIONAL P R O G R A M SURVEILLANCE
Y.C. Chien 1, C.F. Jan 2, H.S. Kuo 3, C.J. Chen 1. 1Graduate Institute of Epidemiology, College of Public Health, 2Department of Family Medicine, College of Medicine, National Taiwan University, Taipei, Taiwan," 3Center for Disease Control, Taiwan Background and Aims: The national hepatitis B virus (HBV) vaccination program was launched in Taiwan in July 1984. During its first two years (July 1984 to June 1986), only newborns to high-risk (HBsAg-seropositive) mothers were vaccinated. The national vaccination program was extended to all newborns after July 1986. Vaccination records of all vaccinated newborns were stored in a computerized database of the Center for Disease Control in Taiwan. This study aimed to evaluate the efficacy of HBV vaccination on the reduction of incidence of hepatocellular carcinoma (HCC) among vaccinated boys and girls in this national program. Methods: We constructed a cohort of 3,855,485 newborns with their HBV vaccination status by data linkage of computerized national HBV vaccination database (from July 1984 to March 2000) with the national household registry profile in Taiwan. We ascertained newly diagnosed hepatocellular carcinoma (HCC) of cohort subjects through data linkage with computerized profiles of national cancer registry. The duration of follow-up for each subject was calculated from the date of birth to the date of the diagnosis of newly developed HCC, the date of death, or the last date when the national cancer registry data were available (June 30, 2004), whichever came first. Cox's proportional hazards regression analysis was used to estimate the relative risk of developing HCC for various status of HBV vaccination.
$99
Results: During the follow-up of 43,134,217 person-years, there were 158 newly diagnosed HCC. The HCC incidence rate per 100,000 person-years was 0.44 and 0.29 for boys and girls, respectively. The HCC incidence rate was 0.74, 0.51 and 0.31 per 100,000 person-years, respectively, for boys having received 1 2 doses, 3 doses and 4 doses of HBV vaccine. The corresponding figures were 0.43, 0.37 and 0.19 per 100,000 personyears, respectively, for girls. Compared with those who received 1 2 doses of HBV vaccine, the efficacy of preventing HCC was 58% for receiving 4 doses of HBV vaccine. Conclusions: The national HBV vaccination program has significantly reduced the incidence of HCC among vaccinees. Vaccinated boys have a higher HCC incidence than vaccinated girls.
•
COMPARISON OF STAGING SYSTEMS FOR HEPATOCELLULAR C A R C I N O M A IN A JAPANESE COHORT
H. Chung 1, M. Kudo 1, S. Takahashi 1, T. Inoue 1, Y. Sakaguchi 1, S. Hagiwara 1, Y. Minami 1, K. Ueshima 1, T. Fukunaga 1, T. Matsunaga2.
1Department of Gastroenterology and Hepatology, Kinki University School of Medicine, Osaka-sayama, Osaka, Japan," :Clinical Laboratory and Medical Informatics, Osaka Medical Center for Cancer and Cardiovascular Diseases, Osaka, Japan Background and Aims: Although various staging systems for hepatocellular carcinoma (HCC) have been developed in recent years, there is no consensus which staging system is best in predicting the survival of HCC patients. The aim of this study is to compare the performance of JIS score, Barcelona Clinic Liver Cancer (BCLC) staging classification and Tokyo score. Methods: A total of 290 consecutive patients with HCC before initial treatment at Kinki University between January 1999 and December 2001 were included. Patients were stratified according to the three staging classifications. The cumulative survival rates were estimated by the Kaplan Meier method, and the differences in survival rates between the stages were analyzed by the log-rank test. The performance of the staging systems were evaluated by calculating the likelihood ratio (LR))~2 test and Akaike Information Criterion (AIC) based on the Cox regression analysis. Results: The cumulative survival rates of 1, 3, 5-year in the three staging systems were 97/89/78%, 97/78/47%, 75/42/25%, 55/13/5%, 25/10/0%, 0/0/0% (JIS score 0 5), 97/89/78%, 91/65/41%, 71/33/18%, 35/6/3%, 32/16/0% (BCLC stage 0, A, B, C, D), 100/92/81%, 98/77/52%, 87/66/44%, 74/35/10%, 56/27/20%, 55/14/0%, 28/14/14%, 22/11/0%, 0/0/0% (Tokyo score 0 8). There were significant differences between all the stages in the JIS score. However, no significant difference was found between stages C and D in the BCLC staging classification, and between all the scores except score 0 and 1, 2 and 3 in the Tokyo score. The value of L R )~2 test was highest and the value of AIC was lowest in the JIS score among the three staging systems for all patients (n 290), radical treatment group (n 208) and non-radical treatment group (n 82). Conclusion: The JIS score provided the best prognostic stratification for our cohort of patients regardless of the radical, non-radical treatment among the three staging systems besides its simplicity to obtain.
~RISK FACTORS AND PATTERNS OF EXTRAHEPATIC METASTASIS OF HEPATOCELLULAR C A R C I N O M A W.J. Chung, H.S. Lee, S.Y. Kim, Y.S. Lee, K.W. Chung, B.K. Jang, K.S. Park, K.B. Cho, J.S. Hwang, S.H. Ahn. Internal medicine/Keimyung
University Hospital, South Korea Background and Aims: The treatment of extrahepatic metastasis of hepatocellular carcinoma is not easy. So, early detection is very important. We evaluated retrospectively the risk factors and patterns of extrahepatic metastasis of hepatocelluar carcinoma at single center.
POSTERS
S 100
Methods: From January 2001 to March 2005, 881 hepatocelluar carcinoma patients were included. We examined aFE CT, bone scan, MRI, or biopsy of suspected site and divided patients into metastasis(+) group and metastasis( ) group at initial diagnosis. We retrospectively evaluated risk factors and patterns of extrahepatic metastasis of metastasis( ) group at initial diagnosis. Statistical analysis was done by chi-square and Mann Whitney's U-test. Results: 804 patients (91.2%) were included in metastasis( ) group at initial diagnosis. Male/female ratio was 633:171. The median age is 59.2 years. The causes of hepatocellular carcinoma were chronic hepatitis B 69.7% (560), chronic hepatitis C 12% (97), combined chronic hepatitis B and C 3 (0.4%), alcohol 10.7% (82), autoimmune 1 (0.1%), unknown origin 7.6% (61). Significant differences were present in the aFP level (87.71ng/ml vs 3028ng/ml, p 0.001), the presence of portal vein thrombosis (4.6% vs 19%, p 0.001), tumor size (3.75 cm vs ll.3cm, p 0.001) and the number of tumor masses (n 1.5 vs 9.0, p 0.001) in metastasis( ) and metastasis(+) group at initial diagnosis. After 1 year follow-up of metastasis( ) patients, extrahepatic metastases were detected in 65 patients (8.1%). Between the metastasis ( ) and (+) groups, significant difference were present in aFP level (75.04 ng/ml vs 332.86 ng/ml, p 0.001), the presence of portal vein thrombosis (6.2% vs 13.7%, p 0.002), tumor size (3.5 cm vs 8.0 cm, p 0.001), and the number of tumor masses (n 1.0 vs 9.0, p 0.001). The sites of metastasis were lung 60.0% (39), regional lymph node 30.8% (20), bone 23.1% (15), perinoenum 12.3% (8), adrenal gland 9.2% (6), brain 1.5% (1), neck node 1.5% (1), spleen 1.5% (1). Conclusions: Serum aFP level, the presence of portal vein thrombosis, size of tumor, and the number of tumor masses can be the risk factors of extrahepatic metastasis of hepatocellular carcinoma. Common sites of extrahepatic metastasis of hepatocellular carcinoma were lung (60%), regional lymph nodes (30.8%) and the bone (23.1%). But, we need more studies about precise cut-off level of each parameters.
•
SUB-CELLULAR EXPRESSION OF Apel/Refl IN HEPATOCELLULAR CARCINOMA AT DIFFERENT STAGES: POSSIBLE PROGNOSTIC SIGNIFICANCE
V. Di Maso 1'2, L.S. Croc~ 1'2'3, C. Avellini 4, G. Tell5, V. Del Mistro 1, S. Intini6, F. Cattin 6, F. Sasso7, C. Tiribelli 1.2.3.1Centro Studi Fegato/Area
Science Park, Trieste, Italy," 2Centro Clinico Studi Fegato/AOU Cattinara, Trieste, Italy," 3Dept. Bioehemistry, University of Trieste, Italy," 4Seienze Biomediehe e Morfologiehe, University of Udine, Italy," 5Seienze e Tecnologie Biomediehe, University of Udine, Italy," 6Seienze Chirurgiehe, University of Udine, Italy," 7Dept. of Surgery, University of Trieste, Italy Background and Alms: Alteration of Redox mechanisms and changes in base repair functions (BER) are associated with chronic liver disease and human hepatocellular carcinoma (HCC). Apel/Refl is involved in cellular redox state and is the major constituent of BER pathway. Apel/Refl sub-cellular localization has been shown to have a possible prognostic significance in breast and colorectal cancer, but no data are available for HCC. The aim of the study was to assess Apel/Refl variable immunophenotypical expression in HCC and surrounding liver cirrhosis (SLC). Methods: 57 consecutive liver surgical specimens of HCC were analyzed by immunohistochemical methods for Apel/Refl. Each sample included HCC tissue and SLC. Nuclear and cytoplasmic Apel/Refl expression was evaluated by a single reader (C.A.) as a percentage of positive cells on 1000 hepatocytes for each slide in both HCC and SLC. The survival rate according to Apel/Refl sub-cellular localization was calculated. Results: Nuclear reactivity for Apel/Refl was comparable in HCC and SLC (80% and 69% respectively) while cytoplasmic reactivity was significantly more frequent in HCC tissue than in SLC (53% vs. 15%, p < 0.04). By stratifying HCC grading according to Edmondson and Steiner criteria, 42% of our patients were GI/G2 (well differentiated HCC) and 58% were G3/G4 (poorly differentiated HCC). Cytoplasmic localization in HCC
tissue was significantly more frequent in G3/G4 than in GI/G2 HCC (71% vs. 39%, p < 0.02). No difference in Ape l/Refl expression was found among HBV, HCV and alcohol related HCC or the CLIP score. Patients with cytoplasmic reactivity for Apel/Refl in HCC tissue showed a shorter survival rate following resection (17• months) than those negative for cytoplasmic localization (34• months) (Cox Sig.0.042). Conclusions: Apel/Refl cytoplasmic localization was: (1) significantly more frequent in HCC than in SLC; (2) associated with a lower degree of differentiation; (3) associated with a survival time of almost half that in cytoplasm negative cases. Collectively, these data suggest a possible role of Apel/Refl sub-cellular localization as a prognostic marker for HCC, with a significant association to neoplastic differentiation grading.
[-~A
siRNA TARGETED AGAINST SRF REDUCES HEPATOCELLULAR CARCINOMA CELL PROLIFERATION, SHOWING ITS POTENTIAL USE IN THE GROWTH INHIBITION OF THIS TYPE OF TUMOUR
R. Farra 1, G. Pozzato 1, B. Dapas 1, F. Agostini 1, B. Scaggiante 2, G. Guarnieri 1, M. Colombo 3, G. Grassi 1. 1Department of Internal
Medicine, University Hospital of Cattinara, Trieste, Italy," 2Department of Biochemistry, Biophysics and Macromolecular Chemistry, University of Trieste, Trieste, Italy," 3Dipartimento di Medicina Interna Universit~ degli Studi di Milano Fondazione Ospedale Maggiore di Milano, Mangiagalli e Regina Elena, Milano, Italy Background and Alms: hepatocellular carcinoma (HCC) is the third leading cause of cancer death, with an estimated 564,000 new cases and almost as many deaths in 2000. Despite therapies, local recurrences are the role, life expectancy is short and there is no clinically useful therapy for the advanced stage of this tumour. Recently, it has been shown that the growth inhibitory effect on HCC of the synthetic acyclic retinoid (ACR) (Run-Xuan et all, Gastroenterology, 2005;128:86 95), is based on the inhibition of FGF-mediated signalling which in turn suppresses the activity of Rho and the serum response factor (SRF) mediated transcription. This observation prompted us to generate an siRNA directed against SRF and test its anti-proliferative effect in cultured HCC cells. Methods: the down-regulation of the target gene has been achieved by siRNA (220 riM) delivered via liposomes to cultured HepG2 cells, undertaken as a HCC cell model. As control, non-treated cells and cells treated by an irrelevant siRNA directed against the luciferase gene (siGL2), were considered. The effects on protein levels were evaluated by western blot. Cell proliferation was measured by cell counting and evaluating BrdU incorporation and propidium iodide intercalation into newly synthesised DNA by flow cytometry. In addition the level of relevant cell cycle genes such as cyclin El, the transcription factor E2F1 and the retinoblastoma protein pRb, were evaluated by western blot. Results: maximal reduction of SRF protein level was achieved two days after transfection and correspond to 60% of control. At the same time point, whereas the protein levels of E2F1 and the hyper-phosphorylated form of pRb were slightly reduced, no variation were observed for cyclin El. The percent of BrdU positive cells, proportional to the amount of proliferating cells, was reduced down to 50% of control. Finally, cell number was reduced down to 50% of controls for at least 3 days after transfection. Conclusions: the specific depletion of SRF, achieved by the selected siRNA, reduces HCC cell growth directly proving the role of SRF in HCC cell proliferation and showing the therapeutic potential of our siRNA in the prevention of HCC development.
I•
EFFECT OF LOW DOSE 5-FLUOROURACIL AND CISPLATIN INTRA-ARTERIAL INFUSION C H E M O T H E R A P Y IN ADVANCED HEPATOCELLULAR C A R C I N O M A WITH DECOMPENSATED CIRRHOSIS
J.Y. Cheong 1, S.W. Cho 1, T.Y. Lim 1, K.M. Lee 1, K.B. Hahm2, J.H. Kim 1.
1Dep. Gastroenterology, 2Genomic Research Center for Gastroenterology, Ajou University School of Medicine, Suwon, South Korea Background and Aims: Advanced hepatocellular carcinoma (HCC) with portal vein thrombosis (PVT) has a poor prognosis. The aim of this study was to evaluate the efficacy and safety of repeated arterial infusion of low dose cisplatin and 5-fluorouracil (FU) in patients who were diagnosed as advanced HCC with decompensated cirrhosis. Methods: Between January 1995 and December 2003, a total of 79 decompensated cirrhotic patients with HCC having PVT were analyzed and divided into 2 groups. Group 1 (n 40) received intra-arterial infusion chemotherapy with cisplatin (10mg for 5 days) and 5-FU (250mg for 5 days) via implanted chemoport, and group 2 (n 39) was managed with only conservative treatment. Results: The two groups were well matched with respect to features relating to prognosis including age, sex, Chil~Pugh class. Although diffuse tumor involvement, main portal vein tumor thrombosis and bi-lobar involvement were more frequent in group 1, the median survival period of group 1 was significantly longer than group 2 (5 months vs. 3 months, P 0.0156). Also, the 1-year survival rate of group 1 (7.5%) was higher than that of group 2 (5.1%) (P 0.0156). If we analyze the patients with Child class B, survival benefits of intra-arterial chemotherapy were more significant (P 0.0077). Conclusions: Intra-arterial chemotherapy consisting of low dose 5-FU and cisplatin achieved favorable result in advanced HCC patients who had decompensated cirrhosis and showed better survival in selected patients. This therapy may be useful as a palliative treatment for HCC patients in decompensated cirrhosis.
I-~
EFFICACY OF HEPATITIS B VIRUS VACCINATION ON THE REDUCTION OF HEPATOCELLULAR C A R C I N O M A INCIDENCE: POST-NATIONAL P R O G R A M SURVEILLANCE
Y.C. Chien 1, C.F. Jan 2, H.S. Kuo 3, C.J. Chen 1. 1Graduate Institute of Epidemiology, College of Public Health, 2Department of Family Medicine, College of Medicine, National Taiwan University, Taipei, Taiwan," 3Center for Disease Control, Taiwan Background and Aims: The national hepatitis B virus (HBV) vaccination program was launched in Taiwan in July 1984. During its first two years (July 1984 to June 1986), only newborns to high-risk (HBsAg-seropositive) mothers were vaccinated. The national vaccination program was extended to all newborns after July 1986. Vaccination records of all vaccinated newborns were stored in a computerized database of the Center for Disease Control in Taiwan. This study aimed to evaluate the efficacy of HBV vaccination on the reduction of incidence of hepatocellular carcinoma (HCC) among vaccinated boys and girls in this national program. Methods: We constructed a cohort of 3,855,485 newborns with their HBV vaccination status by data linkage of computerized national HBV vaccination database (from July 1984 to March 2000) with the national household registry profile in Taiwan. We ascertained newly diagnosed hepatocellular carcinoma (HCC) of cohort subjects through data linkage with computerized profiles of national cancer registry. The duration of follow-up for each subject was calculated from the date of birth to the date of the diagnosis of newly developed HCC, the date of death, or the last date when the national cancer registry data were available (June 30, 2004), whichever came first. Cox's proportional hazards regression analysis was used to estimate the relative risk of developing HCC for various status of HBV vaccination.
$99
Results: During the follow-up of 43,134,217 person-years, there were 158 newly diagnosed HCC. The HCC incidence rate per 100,000 person-years was 0.44 and 0.29 for boys and girls, respectively. The HCC incidence rate was 0.74, 0.51 and 0.31 per 100,000 person-years, respectively, for boys having received 1 2 doses, 3 doses and 4 doses of HBV vaccine. The corresponding figures were 0.43, 0.37 and 0.19 per 100,000 personyears, respectively, for girls. Compared with those who received 1 2 doses of HBV vaccine, the efficacy of preventing HCC was 58% for receiving 4 doses of HBV vaccine. Conclusions: The national HBV vaccination program has significantly reduced the incidence of HCC among vaccinees. Vaccinated boys have a higher HCC incidence than vaccinated girls.
•
COMPARISON OF STAGING SYSTEMS FOR HEPATOCELLULAR C A R C I N O M A IN A JAPANESE COHORT
H. Chung 1, M. Kudo 1, S. Takahashi 1, T. Inoue 1, Y. Sakaguchi 1, S. Hagiwara 1, Y. Minami 1, K. Ueshima 1, T. Fukunaga 1, T. Matsunaga2.
1Department of Gastroenterology and Hepatology, Kinki University School of Medicine, Osaka-sayama, Osaka, Japan," :Clinical Laboratory and Medical Informatics, Osaka Medical Center for Cancer and Cardiovascular Diseases, Osaka, Japan Background and Aims: Although various staging systems for hepatocellular carcinoma (HCC) have been developed in recent years, there is no consensus which staging system is best in predicting the survival of HCC patients. The aim of this study is to compare the performance of JIS score, Barcelona Clinic Liver Cancer (BCLC) staging classification and Tokyo score. Methods: A total of 290 consecutive patients with HCC before initial treatment at Kinki University between January 1999 and December 2001 were included. Patients were stratified according to the three staging classifications. The cumulative survival rates were estimated by the Kaplan Meier method, and the differences in survival rates between the stages were analyzed by the log-rank test. The performance of the staging systems were evaluated by calculating the likelihood ratio (LR))~2 test and Akaike Information Criterion (AIC) based on the Cox regression analysis. Results: The cumulative survival rates of 1, 3, 5-year in the three staging systems were 97/89/78%, 97/78/47%, 75/42/25%, 55/13/5%, 25/10/0%, 0/0/0% (JIS score 0 5), 97/89/78%, 91/65/41%, 71/33/18%, 35/6/3%, 32/16/0% (BCLC stage 0, A, B, C, D), 100/92/81%, 98/77/52%, 87/66/44%, 74/35/10%, 56/27/20%, 55/14/0%, 28/14/14%, 22/11/0%, 0/0/0% (Tokyo score 0 8). There were significant differences between all the stages in the JIS score. However, no significant difference was found between stages C and D in the BCLC staging classification, and between all the scores except score 0 and 1, 2 and 3 in the Tokyo score. The value of L R )~2 test was highest and the value of AIC was lowest in the JIS score among the three staging systems for all patients (n 290), radical treatment group (n 208) and non-radical treatment group (n 82). Conclusion: The JIS score provided the best prognostic stratification for our cohort of patients regardless of the radical, non-radical treatment among the three staging systems besides its simplicity to obtain.
~RISK FACTORS AND PATTERNS OF EXTRAHEPATIC METASTASIS OF HEPATOCELLULAR C A R C I N O M A W.J. Chung, H.S. Lee, S.Y. Kim, Y.S. Lee, K.W. Chung, B.K. Jang, K.S. Park, K.B. Cho, J.S. Hwang, S.H. Ahn. Internal medicine/Keimyung
University Hospital, South Korea Background and Aims: The treatment of extrahepatic metastasis of hepatocellular carcinoma is not easy. So, early detection is very important. We evaluated retrospectively the risk factors and patterns of extrahepatic metastasis of hepatocelluar carcinoma at single center.
POSTERS
S 100
Methods: From January 2001 to March 2005, 881 hepatocelluar carcinoma patients were included. We examined aFE CT, bone scan, MRI, or biopsy of suspected site and divided patients into metastasis(+) group and metastasis( ) group at initial diagnosis. We retrospectively evaluated risk factors and patterns of extrahepatic metastasis of metastasis( ) group at initial diagnosis. Statistical analysis was done by chi-square and Mann Whitney's U-test. Results: 804 patients (91.2%) were included in metastasis( ) group at initial diagnosis. Male/female ratio was 633:171. The median age is 59.2 years. The causes of hepatocellular carcinoma were chronic hepatitis B 69.7% (560), chronic hepatitis C 12% (97), combined chronic hepatitis B and C 3 (0.4%), alcohol 10.7% (82), autoimmune 1 (0.1%), unknown origin 7.6% (61). Significant differences were present in the aFP level (87.71ng/ml vs 3028ng/ml, p 0.001), the presence of portal vein thrombosis (4.6% vs 19%, p 0.001), tumor size (3.75 cm vs ll.3cm, p 0.001) and the number of tumor masses (n 1.5 vs 9.0, p 0.001) in metastasis( ) and metastasis(+) group at initial diagnosis. After 1 year follow-up of metastasis( ) patients, extrahepatic metastases were detected in 65 patients (8.1%). Between the metastasis ( ) and (+) groups, significant difference were present in aFP level (75.04 ng/ml vs 332.86 ng/ml, p 0.001), the presence of portal vein thrombosis (6.2% vs 13.7%, p 0.002), tumor size (3.5 cm vs 8.0 cm, p 0.001), and the number of tumor masses (n 1.0 vs 9.0, p 0.001). The sites of metastasis were lung 60.0% (39), regional lymph node 30.8% (20), bone 23.1% (15), perinoenum 12.3% (8), adrenal gland 9.2% (6), brain 1.5% (1), neck node 1.5% (1), spleen 1.5% (1). Conclusions: Serum aFP level, the presence of portal vein thrombosis, size of tumor, and the number of tumor masses can be the risk factors of extrahepatic metastasis of hepatocellular carcinoma. Common sites of extrahepatic metastasis of hepatocellular carcinoma were lung (60%), regional lymph nodes (30.8%) and the bone (23.1%). But, we need more studies about precise cut-off level of each parameters.
•
SUB-CELLULAR EXPRESSION OF Apel/Refl IN HEPATOCELLULAR CARCINOMA AT DIFFERENT STAGES: POSSIBLE PROGNOSTIC SIGNIFICANCE
V. Di Maso 1'2, L.S. Croc~ 1'2'3, C. Avellini 4, G. Tell5, V. Del Mistro 1, S. Intini6, F. Cattin 6, F. Sasso7, C. Tiribelli 1.2.3.1Centro Studi Fegato/Area
Science Park, Trieste, Italy," 2Centro Clinico Studi Fegato/AOU Cattinara, Trieste, Italy," 3Dept. Bioehemistry, University of Trieste, Italy," 4Seienze Biomediehe e Morfologiehe, University of Udine, Italy," 5Seienze e Tecnologie Biomediehe, University of Udine, Italy," 6Seienze Chirurgiehe, University of Udine, Italy," 7Dept. of Surgery, University of Trieste, Italy Background and Alms: Alteration of Redox mechanisms and changes in base repair functions (BER) are associated with chronic liver disease and human hepatocellular carcinoma (HCC). Apel/Refl is involved in cellular redox state and is the major constituent of BER pathway. Apel/Refl sub-cellular localization has been shown to have a possible prognostic significance in breast and colorectal cancer, but no data are available for HCC. The aim of the study was to assess Apel/Refl variable immunophenotypical expression in HCC and surrounding liver cirrhosis (SLC). Methods: 57 consecutive liver surgical specimens of HCC were analyzed by immunohistochemical methods for Apel/Refl. Each sample included HCC tissue and SLC. Nuclear and cytoplasmic Apel/Refl expression was evaluated by a single reader (C.A.) as a percentage of positive cells on 1000 hepatocytes for each slide in both HCC and SLC. The survival rate according to Apel/Refl sub-cellular localization was calculated. Results: Nuclear reactivity for Apel/Refl was comparable in HCC and SLC (80% and 69% respectively) while cytoplasmic reactivity was significantly more frequent in HCC tissue than in SLC (53% vs. 15%, p < 0.04). By stratifying HCC grading according to Edmondson and Steiner criteria, 42% of our patients were GI/G2 (well differentiated HCC) and 58% were G3/G4 (poorly differentiated HCC). Cytoplasmic localization in HCC
tissue was significantly more frequent in G3/G4 than in GI/G2 HCC (71% vs. 39%, p < 0.02). No difference in Ape l/Refl expression was found among HBV, HCV and alcohol related HCC or the CLIP score. Patients with cytoplasmic reactivity for Apel/Refl in HCC tissue showed a shorter survival rate following resection (17• months) than those negative for cytoplasmic localization (34• months) (Cox Sig.0.042). Conclusions: Apel/Refl cytoplasmic localization was: (1) significantly more frequent in HCC than in SLC; (2) associated with a lower degree of differentiation; (3) associated with a survival time of almost half that in cytoplasm negative cases. Collectively, these data suggest a possible role of Apel/Refl sub-cellular localization as a prognostic marker for HCC, with a significant association to neoplastic differentiation grading.
[-~A
siRNA TARGETED AGAINST SRF REDUCES HEPATOCELLULAR CARCINOMA CELL PROLIFERATION, SHOWING ITS POTENTIAL USE IN THE GROWTH INHIBITION OF THIS TYPE OF TUMOUR
R. Farra 1, G. Pozzato 1, B. Dapas 1, F. Agostini 1, B. Scaggiante 2, G. Guarnieri 1, M. Colombo 3, G. Grassi 1. 1Department of Internal
Medicine, University Hospital of Cattinara, Trieste, Italy," 2Department of Biochemistry, Biophysics and Macromolecular Chemistry, University of Trieste, Trieste, Italy," 3Dipartimento di Medicina Interna Universit~ degli Studi di Milano Fondazione Ospedale Maggiore di Milano, Mangiagalli e Regina Elena, Milano, Italy Background and Alms: hepatocellular carcinoma (HCC) is the third leading cause of cancer death, with an estimated 564,000 new cases and almost as many deaths in 2000. Despite therapies, local recurrences are the role, life expectancy is short and there is no clinically useful therapy for the advanced stage of this tumour. Recently, it has been shown that the growth inhibitory effect on HCC of the synthetic acyclic retinoid (ACR) (Run-Xuan et all, Gastroenterology, 2005;128:86 95), is based on the inhibition of FGF-mediated signalling which in turn suppresses the activity of Rho and the serum response factor (SRF) mediated transcription. This observation prompted us to generate an siRNA directed against SRF and test its anti-proliferative effect in cultured HCC cells. Methods: the down-regulation of the target gene has been achieved by siRNA (220 riM) delivered via liposomes to cultured HepG2 cells, undertaken as a HCC cell model. As control, non-treated cells and cells treated by an irrelevant siRNA directed against the luciferase gene (siGL2), were considered. The effects on protein levels were evaluated by western blot. Cell proliferation was measured by cell counting and evaluating BrdU incorporation and propidium iodide intercalation into newly synthesised DNA by flow cytometry. In addition the level of relevant cell cycle genes such as cyclin El, the transcription factor E2F1 and the retinoblastoma protein pRb, were evaluated by western blot. Results: maximal reduction of SRF protein level was achieved two days after transfection and correspond to 60% of control. At the same time point, whereas the protein levels of E2F1 and the hyper-phosphorylated form of pRb were slightly reduced, no variation were observed for cyclin El. The percent of BrdU positive cells, proportional to the amount of proliferating cells, was reduced down to 50% of control. Finally, cell number was reduced down to 50% of controls for at least 3 days after transfection. Conclusions: the specific depletion of SRF, achieved by the selected siRNA, reduces HCC cell growth directly proving the role of SRF in HCC cell proliferation and showing the therapeutic potential of our siRNA in the prevention of HCC development.