- Email: [email protected]
Bn Rats
2, 37 (48.7%); 3, 6 (7.9%): p27kip1 intensity - 0, 23 (30.3%); 1, 31 (40.8%); 2, 15 (19.7%); 3, 7 (9.2%): Ki67 - negative, 33 (43.4%); positive, 43 (56.6%). The staining intensity of Skp2 and Cks1 showed significant linear correlation (Pearson's R 0.427, p < 0.01). Each staining intensity of Skp2 and Cks1 also showed significant linear correlations with the staining intensity of p27kip1, respectively (Pearson's R 0.314, p < 0.01 for Skp2 and p27kip1, Pearson's R 0.403, p < 0.01 for Cks1 and p27kip1). The positive immunostaining group for Ki67 showed significantly stronger immunostaining intensity for Skp2 (p < 0.01), Cks1 (p < 0.01) and cytoplasmic p27kip1 (p = 0.033). By Kaplan-Meier analyses, the higher IS (p = 0.018) and stronger immunostaining intensity (p < 0.01) of Skp2 and Cks1 were significantly associated with shorter survival for patients with extrahepatic cholangiocarcinoma. Conclusions: Overexpression of Skp2/Cks1 and aberrant localization of p27kip1 into cytoplasm might be useful predictors of prognosis in patients with extrahepatic bile duct cancer.
HCC was observed one-year after TACE. One-year recurrence of HCC was not associated with any independent risk factor studied. Child classification≧B is an independent risk factor of two-year mortality among patients who underwent TACE for HCC.
Combination Therapy of Gemcitabine and Irinotecan in Advanced Biliary Tract Cancer: Phase II Trial Moon Jae Chung, Semi Park, Jeong Youp Park, Seungmin Bang, Seung Woo Park, Si Young Song Background and Aim: Biliary tract cancer (BTC) is often diagnosed at advanced stage and has a poor prognosis. Chemotherapy is palliative treatment option in advanced biliary tract cancer. The aim of this phase II trial was to evaluate the efficacy and safety of the combination therapy of gemcitabine and irinotecan in patients with previously untreated advanced BTC. Patients and Methods: Patients with advanced biliary tract cancer were enrolled in this study. Patients received gemcitabine 1,000mg/m2 (day 1 and 8) and irinotecan 100mg/m2 (day 1 and 8), every 3 weeks. We performed interim analysis of 29 patients who completed the treatment. Results: Among 29 patients eligible for this trial, 24 patients were treated. Nineteen and five patients had gallbladder cancer and peripheral cholangiocarcinoma. A total of 132 cycles of chemotherapy were delivered, with a median of 4.5 cycles of treatment (2-12, 95% CI) per patient. The objective response rate and disease control rate were 16.7 % (4/24 patients) and 70.8% (17/24) in the treated patients. Median progression free survival was 4.4 months (2.4-6.5, 95% CI) and median overall survival was 8.6 months (3.2-14.0, 95% CI). Grade 3/4 toxicities included anemia (20.7% of patients), thrombocytopenia (10.3%), neutropenia (10.3%), AST/ALT increase (13.8%), and emesis (10.3%). Conclusions: Gemcitabine and irinotecan combination chemotherapy had modest efficacy and was well tolerated in patients with advanced BTC.
W1894 Elucidating Gene Regulatory Network of COOH-Terminal Truncated Hbx Associated With Hepatocellular Carcinoma Alfred S. Cheng, Ranxu Zhu, Suki S. Lau, Wing Kit Yip, Yangchao Chen, Vincent W. Wong, Henry Lik-Yuen Chan, Joseph J. Sung Background and Aims: Chronic hepatitis B virus (HBV) infection is the major cause of hepatocellular carcinoma (HCC) in Southeast Asia. Although the X protein of HBV (HBx) is highly implicated in hepatocarcinogenesis, the molecular mechanisms are not fully explored. HBx not only activates cytoplasmic signaling cascades but also physically interacts with DNA-bound transcription factors to directly regulate gene transcription in the nucleus. Recent studies have shown that COOH-terminal truncated HBx loses the growth-suppressive effect of full-length HBx and enhances the transformation ability. We aimed to elucidate the direct target gene promoters and interacting transcription factor partners that constitute the gene regulatory network of COOH-terminal truncated HBx in human hepatocytes. Methods: COOH-terminally truncated HBx (deletion of 14- or 35-amino acid) was amplified from serum of an HCC patient infected with HBV subgenotype Ce carrying the linked-point mutation (L130M and V131I), cloned into lentiviral vector and over-expressed in human non-tumorigenic hepatocyte MIHA cells. Chromatin immunoprecipitation microarray (or ChIP-chip) was performed in conjunction with human promoter arrays containing ~17,000 best defined human transcripts to identify the target promoters of truncated HBx. Transcription factor binding sites in the bound promoter regions were investigated by MATCH software using the TRANSFAC database. Results: Over-expression of truncated HBx abrogated the growth-suppressive effect of full-length HBx. ChIP-chip analysis identified 1-2% (~250) of the interrogated promoters that were likely regulated by truncated HBx in human hepatocytes. Consistent with the reported function of HBx in cell invasion, genes involved in cell adhesion, motility and migration were significantly enriched. Notably, a high proportion of novel target genes possessed Gene Ontology annotations for transcriptional regulation e.g. the oncogenic transcription factors PAX9 and PITX2. Bioinformatics analysis of HBx-bound promoter regions further revealed over-representation of transcription factor binding sites of STAT, NF-kappaB, E2F, etc that have been shown to promote hepatocarcinogenesis. Conclusions: The fact that many truncated HBx target genes are also transcription factors/ regulators implies a chain reaction, which may explain the cross-activation of multiple pathways by HBx. Further characterization of the functional interaction between truncated HBx and the putative transcription factor partners in gene regulation will greatly enrich our understanding of HBV-induced hepatocarcinogenesis and provide potential therapeutic targets for treatment of HCC.
W1897 Visceral and Somatic Pain Reporting in Aged Fisher 344 (F-344), Brown Norway (Bn) and F344/Bn Rats Aaron S. Chaloner, Beverley Greenwood-Van Meerveld Visceral and somatic pain reporting are abnormal in patients with functional bowel disorders, and changes in pain perception have been implicated in age-related pathologies. The goal of our study was to assess visceral and somatic perception in young adult rats and then investigate whether pain perception is altered with aging in the same rodent models. Methods: Experiments were performed in young (6 months) and old (26 months) male rats in three strains available from the National Institute of Aging (Brown Norway (BN), Fisher 344 (F344), and F344-BN). In freely moving rats, we evaluated visceral sensitivity via measurements of the visceromotor response (VMR) to graded colorectal distension (CRD) before and following primary afferent sensitization with intracolonic infusion of dilute acetic acid (0.6%). Somatic pain thresholds were assessed as the minimal force required to elicit hind paw withdrawal in response to a mechanical stimulus. Results: In BN and F-344 rats, prior to colonic sensitization, we observed a linear increase in the VMR to graded pressures (20-60 mmHg) of isobaric CRD that was unaffected by the age of the rats. However, in the F344-BN the number of abdominal contractions induced by the highest distension pressure (60 mmHg) was significantly lower in older rat compared to younger animals (9.8±1.1 vs. 14.7±2.0, P<0.01). Following colonic sensitization, both young and old F-344/BN rats exhibited a significant increase in colonic sensitivity as demonstrated by an exaggerated VMR to CRD. However, in F-344 colonic hypersensitivity which was evident in young rats at all distension pressures, was only significant at the 40 mmHg distension pressure in older animals (11.3±1.5 vs. 17.6±1.6, P <0.05). The BN rat strain was resistant to colonic sensitization except at the highest distension pressure (9.2±1.2 vs. 14.5±1.5, P <0.01) in older animals. Somatic pain thresholds were age-and strain-dependent (BN: young=55.7±10.9, old=88.9±7.3, P<0.05; F344: young=68.5±1.7, old=53.3±1.7, P<0.001; F344-BN: young=89.3±9.9, old=101.6±6.9, P>0.05). Summary: In three different stains of rat, used frequently to investigate the effect of aging, the data demonstrate that age- and strain-related alterations exist in visceral sensitivity and somatic pain reporting. Conclusion: Our result suggest that the underlying neural circuitry that regulates visceral and somatic pain behaviors may explain unique phenotypic differences in pain perception observed with aging in 3 rodent strains.
W1895 Prognosis of Hepatocellular Carcinoma Treated by Transcatheter Arterial Chemoembolization: Risk Factors for One-Year Recurrence and Two-Year Mortality Michitaka Matsuda, Fumio Omata, Takeshi Setoyama, Shoko Suzuki, Masayo Uemura, Naoki Ishii, Yusuke Iizuka, Katsuyuki Fukuda, Yoshiyuki Fujita BACKGROUND & AIMS: Transcatheter arterial chemoembolizaion (TACE) is a therapy of last resort for patients with hepatocellular carcinoma (HCC) for whom liver transplant, resection, or ablation is not possible. Due to its end-stage severity, it is important to know the prognosis of patients initially treated by TACE. While there are some survival analyses, few studies report both mortality and recurrence rates. In this study, we depict the oneyear recurrence and two-year survival rate and determine their independent risk factors. METHODS: We conducted a retrospective cohort study in 43 consecutive patients with HCC undergoing epirubicin-based TACE as initial therapy. Any patient with hepatic or portal vein invasion was excluded from the analysis. None of the patient had lymph node or distant metastasis. For time-to-event analysis of recurrence, 11 patients with incomplete initial TACE were excluded. Adequacy of TACE and tumor recurrence was confirmed by contrast CAT scan every 3 months. We calculated Kaplan-Meier estimates and performed multiple regression analysis using Cox-proportional hazard model. RESULTS: A total of 43 patients (male=60%), all with liver cirrhosis, underwent TACE as initial therapy for HCC from April 2004 to March 2009. 32 (74%) cases and 4 (9%) cases were infected with hepatitis C and hepatitis B, respectively. Mean age (SD), mean platelet count (SD) and mean albumin (SD) were 69 (7.8), 12.8 x104 (6x104) and 3.6 (0.6), respectively. 19 (44%) patients had a single HCC. The median [range] of maximum HCC diameter was 2.7 [0.519] cm. 34 (79%) patients were classified Child A and 9 (21%) were Child B. Kaplan-Meier estimates revealed that one-year recurrence rate [95% CI] and two-year survival rate [95% CI] were 58% [42-75%] and 73% [54-85%], respectively. Bivariate analysis revealed that hazard ratio (HR) [95% CI] of maximum HCC diameter was 1.5 [1.04-2.0] for one-year recurrence and that HR [95%CI] of non-Child A and maximum HCC diameter was 4.6 [1.2-17.4] and 1.2 [1.05-1.5], respectively for two-year mortality. Adjusted HR [95% CI] of age, multiplicity of tumor, maximum tumor size and non-Child A for one-year recurrence was 1.0 [0.96-1.1], 0.74 [0.25-2.2], 1.4 [0.97-2.1] and 1.99 [0.48-8.2], respectively. For two-year survival, adjusted HR [95% CI] of these factors for was 1.1 [0.99-1.3], 0.86 [0.193.9], 1.1 [0.0-1.4] and 9 [1.4-57], respectively. CONCLUSIONS: Substantial recurrence of
W1898 The Effects of Hormone Replacement on Oxidative, Inflammatory and Apoptotic Factors in Aged Rat Colon Smooth Muscle Patricia Pascua, Cristina Camello-Almaraz, Pedro J. Camello, Francisco E. Martín-Cano, E. Vara, Jesus A. Tresguerres, Maria J. Pozo BACKGROUND: There are increasing evidences that aging is associated to oxidative damage, inflammation and apoptosis in different cell types. However there is no information regarding these mechanisms in aged colon. We have studied age-related changes in the inflammatory factors cyclooxygenase-2 (COX2) and nuclear factor kappa-beta (NFκB), the apoptotic enzyme caspase-3 and the oxidative status (H2O2 content and antioxidant enzymes) in rat colonic smooth muscle, and the influence of chronic hormone treatments on them. METHODS: Old male and female Wistar rats (22 months) were used in the study. Two month old animals were used as control. Rats were treated for 10 weeks with the hormone of election: old male and female rats were treated with melatonin (MEL), old male with growth hormone (GH), a group of female rats were castrated at 12 months of age and treated with estrogens (EOS). All aged animals were sacrificed at 24 months by decapitation. After sacrifice the colon was extracted, the mucosa was removed and the smooth muscle was homogenizated. Expression of COX2, caspase3 and NFκB were determined by Western blott. H2O2 content, catalase and glutation peroxidase (GPX) activity were determined by colorimetric kits in homogenizated samples. RESULTS: Aging of colon correlated with an increase in H2O2 levels, when compared to young animals, in both proximal and distal segments, as the result of the decrease in the activity of catalase and GPx in female and
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AGA Abstracts
AGA Abstracts
W1896
HCC was observed one-year after TACE. One-year recurrence of HCC was not associated with any independent risk factor studied. Child classification≧B is an independent risk factor of two-year mortality among patients who underwent TACE for HCC.
Combination Therapy of Gemcitabine and Irinotecan in Advanced Biliary Tract Cancer: Phase II Trial Moon Jae Chung, Semi Park, Jeong Youp Park, Seungmin Bang, Seung Woo Park, Si Young Song Background and Aim: Biliary tract cancer (BTC) is often diagnosed at advanced stage and has a poor prognosis. Chemotherapy is palliative treatment option in advanced biliary tract cancer. The aim of this phase II trial was to evaluate the efficacy and safety of the combination therapy of gemcitabine and irinotecan in patients with previously untreated advanced BTC. Patients and Methods: Patients with advanced biliary tract cancer were enrolled in this study. Patients received gemcitabine 1,000mg/m2 (day 1 and 8) and irinotecan 100mg/m2 (day 1 and 8), every 3 weeks. We performed interim analysis of 29 patients who completed the treatment. Results: Among 29 patients eligible for this trial, 24 patients were treated. Nineteen and five patients had gallbladder cancer and peripheral cholangiocarcinoma. A total of 132 cycles of chemotherapy were delivered, with a median of 4.5 cycles of treatment (2-12, 95% CI) per patient. The objective response rate and disease control rate were 16.7 % (4/24 patients) and 70.8% (17/24) in the treated patients. Median progression free survival was 4.4 months (2.4-6.5, 95% CI) and median overall survival was 8.6 months (3.2-14.0, 95% CI). Grade 3/4 toxicities included anemia (20.7% of patients), thrombocytopenia (10.3%), neutropenia (10.3%), AST/ALT increase (13.8%), and emesis (10.3%). Conclusions: Gemcitabine and irinotecan combination chemotherapy had modest efficacy and was well tolerated in patients with advanced BTC.
W1894 Elucidating Gene Regulatory Network of COOH-Terminal Truncated Hbx Associated With Hepatocellular Carcinoma Alfred S. Cheng, Ranxu Zhu, Suki S. Lau, Wing Kit Yip, Yangchao Chen, Vincent W. Wong, Henry Lik-Yuen Chan, Joseph J. Sung Background and Aims: Chronic hepatitis B virus (HBV) infection is the major cause of hepatocellular carcinoma (HCC) in Southeast Asia. Although the X protein of HBV (HBx) is highly implicated in hepatocarcinogenesis, the molecular mechanisms are not fully explored. HBx not only activates cytoplasmic signaling cascades but also physically interacts with DNA-bound transcription factors to directly regulate gene transcription in the nucleus. Recent studies have shown that COOH-terminal truncated HBx loses the growth-suppressive effect of full-length HBx and enhances the transformation ability. We aimed to elucidate the direct target gene promoters and interacting transcription factor partners that constitute the gene regulatory network of COOH-terminal truncated HBx in human hepatocytes. Methods: COOH-terminally truncated HBx (deletion of 14- or 35-amino acid) was amplified from serum of an HCC patient infected with HBV subgenotype Ce carrying the linked-point mutation (L130M and V131I), cloned into lentiviral vector and over-expressed in human non-tumorigenic hepatocyte MIHA cells. Chromatin immunoprecipitation microarray (or ChIP-chip) was performed in conjunction with human promoter arrays containing ~17,000 best defined human transcripts to identify the target promoters of truncated HBx. Transcription factor binding sites in the bound promoter regions were investigated by MATCH software using the TRANSFAC database. Results: Over-expression of truncated HBx abrogated the growth-suppressive effect of full-length HBx. ChIP-chip analysis identified 1-2% (~250) of the interrogated promoters that were likely regulated by truncated HBx in human hepatocytes. Consistent with the reported function of HBx in cell invasion, genes involved in cell adhesion, motility and migration were significantly enriched. Notably, a high proportion of novel target genes possessed Gene Ontology annotations for transcriptional regulation e.g. the oncogenic transcription factors PAX9 and PITX2. Bioinformatics analysis of HBx-bound promoter regions further revealed over-representation of transcription factor binding sites of STAT, NF-kappaB, E2F, etc that have been shown to promote hepatocarcinogenesis. Conclusions: The fact that many truncated HBx target genes are also transcription factors/ regulators implies a chain reaction, which may explain the cross-activation of multiple pathways by HBx. Further characterization of the functional interaction between truncated HBx and the putative transcription factor partners in gene regulation will greatly enrich our understanding of HBV-induced hepatocarcinogenesis and provide potential therapeutic targets for treatment of HCC.
W1897 Visceral and Somatic Pain Reporting in Aged Fisher 344 (F-344), Brown Norway (Bn) and F344/Bn Rats Aaron S. Chaloner, Beverley Greenwood-Van Meerveld Visceral and somatic pain reporting are abnormal in patients with functional bowel disorders, and changes in pain perception have been implicated in age-related pathologies. The goal of our study was to assess visceral and somatic perception in young adult rats and then investigate whether pain perception is altered with aging in the same rodent models. Methods: Experiments were performed in young (6 months) and old (26 months) male rats in three strains available from the National Institute of Aging (Brown Norway (BN), Fisher 344 (F344), and F344-BN). In freely moving rats, we evaluated visceral sensitivity via measurements of the visceromotor response (VMR) to graded colorectal distension (CRD) before and following primary afferent sensitization with intracolonic infusion of dilute acetic acid (0.6%). Somatic pain thresholds were assessed as the minimal force required to elicit hind paw withdrawal in response to a mechanical stimulus. Results: In BN and F-344 rats, prior to colonic sensitization, we observed a linear increase in the VMR to graded pressures (20-60 mmHg) of isobaric CRD that was unaffected by the age of the rats. However, in the F344-BN the number of abdominal contractions induced by the highest distension pressure (60 mmHg) was significantly lower in older rat compared to younger animals (9.8±1.1 vs. 14.7±2.0, P<0.01). Following colonic sensitization, both young and old F-344/BN rats exhibited a significant increase in colonic sensitivity as demonstrated by an exaggerated VMR to CRD. However, in F-344 colonic hypersensitivity which was evident in young rats at all distension pressures, was only significant at the 40 mmHg distension pressure in older animals (11.3±1.5 vs. 17.6±1.6, P <0.05). The BN rat strain was resistant to colonic sensitization except at the highest distension pressure (9.2±1.2 vs. 14.5±1.5, P <0.01) in older animals. Somatic pain thresholds were age-and strain-dependent (BN: young=55.7±10.9, old=88.9±7.3, P<0.05; F344: young=68.5±1.7, old=53.3±1.7, P<0.001; F344-BN: young=89.3±9.9, old=101.6±6.9, P>0.05). Summary: In three different stains of rat, used frequently to investigate the effect of aging, the data demonstrate that age- and strain-related alterations exist in visceral sensitivity and somatic pain reporting. Conclusion: Our result suggest that the underlying neural circuitry that regulates visceral and somatic pain behaviors may explain unique phenotypic differences in pain perception observed with aging in 3 rodent strains.
W1895 Prognosis of Hepatocellular Carcinoma Treated by Transcatheter Arterial Chemoembolization: Risk Factors for One-Year Recurrence and Two-Year Mortality Michitaka Matsuda, Fumio Omata, Takeshi Setoyama, Shoko Suzuki, Masayo Uemura, Naoki Ishii, Yusuke Iizuka, Katsuyuki Fukuda, Yoshiyuki Fujita BACKGROUND & AIMS: Transcatheter arterial chemoembolizaion (TACE) is a therapy of last resort for patients with hepatocellular carcinoma (HCC) for whom liver transplant, resection, or ablation is not possible. Due to its end-stage severity, it is important to know the prognosis of patients initially treated by TACE. While there are some survival analyses, few studies report both mortality and recurrence rates. In this study, we depict the oneyear recurrence and two-year survival rate and determine their independent risk factors. METHODS: We conducted a retrospective cohort study in 43 consecutive patients with HCC undergoing epirubicin-based TACE as initial therapy. Any patient with hepatic or portal vein invasion was excluded from the analysis. None of the patient had lymph node or distant metastasis. For time-to-event analysis of recurrence, 11 patients with incomplete initial TACE were excluded. Adequacy of TACE and tumor recurrence was confirmed by contrast CAT scan every 3 months. We calculated Kaplan-Meier estimates and performed multiple regression analysis using Cox-proportional hazard model. RESULTS: A total of 43 patients (male=60%), all with liver cirrhosis, underwent TACE as initial therapy for HCC from April 2004 to March 2009. 32 (74%) cases and 4 (9%) cases were infected with hepatitis C and hepatitis B, respectively. Mean age (SD), mean platelet count (SD) and mean albumin (SD) were 69 (7.8), 12.8 x104 (6x104) and 3.6 (0.6), respectively. 19 (44%) patients had a single HCC. The median [range] of maximum HCC diameter was 2.7 [0.519] cm. 34 (79%) patients were classified Child A and 9 (21%) were Child B. Kaplan-Meier estimates revealed that one-year recurrence rate [95% CI] and two-year survival rate [95% CI] were 58% [42-75%] and 73% [54-85%], respectively. Bivariate analysis revealed that hazard ratio (HR) [95% CI] of maximum HCC diameter was 1.5 [1.04-2.0] for one-year recurrence and that HR [95%CI] of non-Child A and maximum HCC diameter was 4.6 [1.2-17.4] and 1.2 [1.05-1.5], respectively for two-year mortality. Adjusted HR [95% CI] of age, multiplicity of tumor, maximum tumor size and non-Child A for one-year recurrence was 1.0 [0.96-1.1], 0.74 [0.25-2.2], 1.4 [0.97-2.1] and 1.99 [0.48-8.2], respectively. For two-year survival, adjusted HR [95% CI] of these factors for was 1.1 [0.99-1.3], 0.86 [0.193.9], 1.1 [0.0-1.4] and 9 [1.4-57], respectively. CONCLUSIONS: Substantial recurrence of
W1898 The Effects of Hormone Replacement on Oxidative, Inflammatory and Apoptotic Factors in Aged Rat Colon Smooth Muscle Patricia Pascua, Cristina Camello-Almaraz, Pedro J. Camello, Francisco E. Martín-Cano, E. Vara, Jesus A. Tresguerres, Maria J. Pozo BACKGROUND: There are increasing evidences that aging is associated to oxidative damage, inflammation and apoptosis in different cell types. However there is no information regarding these mechanisms in aged colon. We have studied age-related changes in the inflammatory factors cyclooxygenase-2 (COX2) and nuclear factor kappa-beta (NFκB), the apoptotic enzyme caspase-3 and the oxidative status (H2O2 content and antioxidant enzymes) in rat colonic smooth muscle, and the influence of chronic hormone treatments on them. METHODS: Old male and female Wistar rats (22 months) were used in the study. Two month old animals were used as control. Rats were treated for 10 weeks with the hormone of election: old male and female rats were treated with melatonin (MEL), old male with growth hormone (GH), a group of female rats were castrated at 12 months of age and treated with estrogens (EOS). All aged animals were sacrificed at 24 months by decapitation. After sacrifice the colon was extracted, the mucosa was removed and the smooth muscle was homogenizated. Expression of COX2, caspase3 and NFκB were determined by Western blott. H2O2 content, catalase and glutation peroxidase (GPX) activity were determined by colorimetric kits in homogenizated samples. RESULTS: Aging of colon correlated with an increase in H2O2 levels, when compared to young animals, in both proximal and distal segments, as the result of the decrease in the activity of catalase and GPx in female and
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AGA Abstracts
AGA Abstracts
W1896