- Email: [email protected]
Prevalence of lewis phenotype (le) in patients suffering from pancreatic diseases
examined by thymidine incorporation and caspase-3 activity. 2) in vwo: A total of 5 x 10~ OE-33 cells were injected subcutaneously into bilateral sides of female BALB/e nude mice Mice were orally given with either vehicle or troglitazone (50, 500 mg/kg/day) for 6 weeks. Tumor size was measured, and proliferation and apoptosis were examined by immunohistochemical staining for Ki-67 and ss-DNA and expressed as labeling index (LI) defined as percentage of positive cells in 5 randomized fields. Result: OE-33 expressed PPAR~' mRNA and protain. Troglitazone significantly inhibited proliferation and induced apoptosis of OE33 cells in a dose-dependant manner. Oral administration of troglitazone did not afl'ect tumor size, but significantly decreased t(i-67 LI fi'om 70,7 -+ 2.5 in controls to 48.9 -+ 1.9 in mice with 50 mg/kg of troglitazone and 42.4 _+ 1.4 in mice with 500 mg/kg of trughiazone (p<0.0001). Trogfitazone siginficantly increased number of apoptotic cells (p<0.05). Conclusion: Troglitazone inhibited proliferation and induced apoptosis of OE33 cells both in vitro and in vivo. These findings suggest that PPAR'y may be a therapeutic target tor human Barrett's adenocarcinoma of the esophagus.
Wl115 Prevalence of Lewis Phenotype (Le) in Patients Suffering from Pancreatic Diseases Luca Fmlloni, Barbara Ca/om, Barbara Ferri, Elena Coato, Sabrina Bmnelfi, Alessandro Canton< Sihqa Can'ara, Laura Bernardoni, Giorgw Cavallini Background: ]qae earbmhydrate antigen CA 1%9, the 2-3 sialylated Lewis a antigen, is a seI~sib/e and specibc marker of the pancreatic carcinoma. It has been shown that m pancreatic cancer the expression of the Le antigen on epflhrocytes could be diminnished. In these patients is therefore increased the prevalence of the Lewis phenothype Le(a-b-). Them are few papers in the literature regarding the prevalence of the phenothype Le in patients affected by pancreatic diseases, Aim: Ain't ot this study was therefore, to evaluate the prevalence of the loewis phenotype I.e(a-b-) and the serum levels of CA 19-9 in relation to Lewis phenotype. Patients and Methods: We studied 57 consecutive patients (35 males, 22 females, mean age 53 -+ 15). 22 patients (39%) wem afl~cted by acute recurrent pancreatitis (ARP); 25 (44%) by chromc pancrcatitis (CP), and 10 (17%) by pancreatic carcinoma (PC) in c o m p a ~ n with 200 healthy controls subjects representing the Italian population. In all these patients we determined the serum level of the CA 19-9 and the Lewis phenothy~ on the erythrocytes. Results: 22 patients (39%) were of the Lewis phenotype Le(a-b-), I1 (19%) Le(a + b-), 22 (39%) Le(a-b + ) and 2 (3%) Lewis(a + b + ).The frequency of the phenothype Le(a-b-) was significantly increased (p<0.00001) compared with 200 healthy' controls. In patients affected by acute recurrent pancreatitis that phenotype was morn frequent (55%) than in patients affected by CP (28%) or by PC(30%) (Tab.).The serum levels of the CA 19-9 were not statistically diff':rent among Le phenotypes. Inside every single disease (AP, CP, and PC), we did not hnd statistlcafiy significant correlation between serum levels of CA 19-9 and the difterent phenots,~s Le. Conclusions: The frequency of phenothype Le(a-b-) is increased m a consecutive series of patients suffering fi'om pancreatic diseases compared with the general population, particularly' in ARP group. The serum levels of the CA 19-9 are not significantly different among phenotypes Le.
W1250 Oxidative Strew-Related Gene Po|ymorphisms and Susceptibility to Barrett's Esophagus Elena Piazuelo, Maria Asuncion Garcia-Gonzalez, Mark Strunk, Javier Alcedo, Juan Arenas, Angel Eerrandez, Pilaf Jimenez, Angel Lanas Both experimental and human studies have demonstrated that free radicals are involved m the ethiopathogenesis of Barretfs esophagus (BE). Antioxidant enz}anes such as superoxide dismutase and catalase plays a pivotal role in protecting esophageal mucosa from the deleterious effects of ROS. Polymorphic variants of the genes encoding these enzymes aftectnig their activity or expression may result in predisposition to Barrett's esophagus. Objective: to determine whether Ala16Val polymorphism in the manganese superoxide dismutase (MnSOD) gene and -262 T/C in the catalase gene are associated with susceptibility to Barmtt's esophagus. Methods: 107 unrelated Spanish Caucasian patients with BE and 286 ethnically matched healthy controls (HC) were included. The diagnosis of BE was established on the basis of endoscopical and histopathological criteria, Genomic DNA was extracted from peripheral blood of cases and controls. Ala 16Val MnSOD and -262T/C catalase gene polymorphisms were determined by RFLP-PCR. Clinical and epideminlogical variables were also collected by structured interview. Statistical analysis was perfbrmed by chi-sqnare test. Results: There were no differences in carriage, genotype, and allele frequencies of Ala16Val MnSOD gene polymorphism between BE patients and HC. However, the dist.qbntion of catalase genotypes was different between BE (CC: 46.2%; CT: 45.3%; TT: 8.5%) and controls (CC: 59.7%; CT: 36.8%; TT: 3.9%) p = 0.03, and carriers of the T allele were more frequent in the group of patients with BE (53.8 %) than in the control group (40.7%).; p = 0.02; R.R, = 1.69 (I.C. = 1.08-2.65). Conclusion: These data suggest that polymorphic variant 262T of catalase gene may be implicated in the susceptibility to Barrett's esophagus.
Prevalence of phenotype Le in different type of pancreatic diseases (PD) Phenotype Le
ARP
a.l~ ~.
12(55%) 6(27%) 4(16%)
a+b*
0,
CP
7(28%) 12(48%) 4(16%) 2(8%)
PC
3(30%) 4(40%} 3(30% 0
All PO
Population
22(39%) 22(39%) 11(19%)
9(4.5%) 147(73.5%) 44(22%)
2(3%) . . . .
0
Wll16 Associated Diseases With Antoimmune Pancreatitis Temmi Kamisawa, Naoto Egawa, Masakatsu Matsukawa Background/Aim: Autoimmune pancreatitis (ALP)sometimes accompanies with other autoimmune diseases or diabetes mellitus (DM). We examined associated diseases and disorders of pancreatic endocrine and exocrine tunction, and function of salivaD" glands in patients with AlP, and transition of these fl.mctions before and after steroid therapy,. Patients: Nineteen patients were diagnosed as AIP according to the following criteria; swelling of the pancreas, irregularly narrowing of the main pancreatic duct, hypergammagloburinemia or autoantibodies, and histologic findings. Steroid therapy, was ettectwe in all 9 patients treated. Methods: We examined blood glucose in hunger, 75-g oral glucose tolerance test, glycosykated hemoglobin values (HbAlc), or urinary C-peptide for pancreatic endocrine function in all patients, and periormed BT-PABA test or secretin test tbr pancreatie exocrine function in 8 patients. For salivary gland function, we measured concentrations of Na + (reflecting function of reabsorption)and beta2 microglobulin (reflecting degree of inflammatinn) in saliva and examined salivary gland scintigraphy with 99mTc in 8 patients. Results: Associated autoimmune diseases were retroperitoneal fibrosis (2 cases), rheumatoid arthritis (2), selemsing cholangitis (2), S/ogren's syndrome (1) and Sarcoidosis (1). Age of patients with these diseases (average 7 0 0 yr.) was older than 65.2 yr. of the others. Eight patients had DM. Seven patients were diagnosed as DM at the same time as AIP, and the other showed exacerbation of DM with AIP. Pancreatic exocrine dysfunction was detected in 7 of 8 cases examined After steroid therapy,, DM improved in four patients, whose pancreatic exocrine fimction also improved. Concentration of Na + and beta2 micmglobulin in saliva of patients with AIP was 22.5+/-9.1 mBoe4 (mean+/-SD) and Z65+/-1.27 mg/dl, which was significantly higher than 13.6 +/-8.4 mE@ and 1.02 +/-0.57 mg/l in a control group respectively (p<0 05). In parotid gland scintigraphy, ratio of cumulative peak count to injected radioisotope, and ratio of washout in patients with AlP was 19.9+/-11.8 % and 51.7+/-32.0 %, significantly lower than 35.3+/-84 % and 8 7 3 + / - 9 . 1 % in a control group, respectively (p<001 ) Impaired salivary gland tunction also improved after steroid therapy. Conclusions: AIP is sometimes associated with mufiifocal fibrusclerosis. Functions of pancreatic endocrine and exocrine, and sahvaD' gland are sometimes impaired, but often improve by steroid fherapy
W1251 Acid and Bile Exposure Cause Additive MAPK Activation and Proliferation in a Barrett's Adenocarcinoma Cell Line George A. Sarosi Jr,, Christie Lopez-Guzman, Stuart Spechier, Rhonda Souza Patients with Barrett's esophagus (BE) have significantly more acid and bile reflux than patients who have gastroesophageal reflux without BE. Exposure to either acid or bile salts alone has been shown to increase profiteration in explants of BE, and in a Barrett's-associated adenocarcinoma cell line (SEG-1). Recent studies suggest that acid and bile indMdually increase proliferation in BE by activating mitogen-activated protein kinases (~,~PKs) such as ERK and p38. Although it has been reported that the combination of acid and bile salts together decrease proliferation in BE, this finding seems counterintuitive. We hypothesized that bile and acid exposure together should have additive effects in activating MAPK signaling and stimulating proliferation in BE, and we tested our hypothesis in SEG-1 ceils. Methods: SEG-1 cells were grown using standard tissue culture techniques. Cells were exposed to the conjugated bile salt glyco-chenodeoxycholic acid (GCDA) at pH 7.4 (bile alone) or at pH 6.0 (bile and acid exposure) for 20 minutes to model reflux episodes. MAPK actMties were measured by immuno-blotting kinase assays. Proliferation was measured by an MTT incorporation assay, Results: Exposure of SEG-1 cells to bile alone over the physiologically relevant range of 50-500 tiM GCDA produced rapid, dose-dependent activation of both the ERK and p38 MAPKs. Exposure to bile and acid produced marked (2 to 4 fold) increases in both ERK and p38 activation beyond that seen with either acid or bile salt exposure alone. Furthermore, exposure to bile and acid produced an additive increase in cell number at 24 hours when compared to either bile salt or acid exposure alone (Table 1)(*, P< 0.05 vs 2001*M GCDA pH 7.4)Conchisions: Combined acid and bile salt exposure produces significant increases in MAPK activation and proliferation in Barmtt's adenocarcinoma cells, beyond that seen with either acid or bile salts alone. These data do not support the published contention that the combined reflux of acid and bile decreases proliferation in Barrett's esophagus.
W1249
Treatme~ Condlton Control
500pM GCDA pH 6.0 100~-2 1247 1216 1176 141 ~5' 132~7 Data expressedas cell number as a percentof ~n~I,,SEM (*,P<0,05 vs 200pM GCDA pH 7,4)
Ligand for Peroxisome Proliferator-Activated Receptor-[' Inhibits Proliferation and Induces Apoptosis of Barrett's Adenocarcinoma Both In Vitro and In Vivo Tsuyoshi Hayakawa, Yasuhiru Fujiwara, Takashi Takashima, Harunori Fujita, Masaki Hamaguchi, Masatsngn Shiba, Kazunari Tominaga, Toshio Watanabe, Nobuhide Oshitani, Kazuhide Higuchb Takayuki Matsumoto, Tetsuo Arakawa
pH 6.0 No Bile
200pM GCDA pH L4
50pM GCDA pH6.0
200pM GCDA pH 6.0
Pemxisume proliferator-activated receptor gamma (PPARy), a member of the nuclear hormone receptor SUlxrfamily, is nivolved in suppression of grmvtb of several tumors. Previous our studies showed that PPAR"/ligands inhibited growth and proliferation of Barrett's adenocarcinuma call lines (TE-7) through induction of G1 cell cycle arrest and apoptosis. The aim of this stud}, was to examine whether PPARy ligand (troglitazone) affects proliferation and apoptosis of Barmtt's adenocaminoma in vivo. Method: 1) in vitro: Human Barrett's adenocarcinoma cells (OE-33) were used. PPARy expression was determined by RT-PCR and western blotting. Efl~cts of troglitazone on proliferation and apoptosis of OE-33 were
A-633
AGA
Abstracts
Wl115 Prevalence of Lewis Phenotype (Le) in Patients Suffering from Pancreatic Diseases Luca Fmlloni, Barbara Ca/om, Barbara Ferri, Elena Coato, Sabrina Bmnelfi, Alessandro Canton< Sihqa Can'ara, Laura Bernardoni, Giorgw Cavallini Background: ]qae earbmhydrate antigen CA 1%9, the 2-3 sialylated Lewis a antigen, is a seI~sib/e and specibc marker of the pancreatic carcinoma. It has been shown that m pancreatic cancer the expression of the Le antigen on epflhrocytes could be diminnished. In these patients is therefore increased the prevalence of the Lewis phenothype Le(a-b-). Them are few papers in the literature regarding the prevalence of the phenothype Le in patients affected by pancreatic diseases, Aim: Ain't ot this study was therefore, to evaluate the prevalence of the loewis phenotype I.e(a-b-) and the serum levels of CA 19-9 in relation to Lewis phenotype. Patients and Methods: We studied 57 consecutive patients (35 males, 22 females, mean age 53 -+ 15). 22 patients (39%) wem afl~cted by acute recurrent pancreatitis (ARP); 25 (44%) by chromc pancrcatitis (CP), and 10 (17%) by pancreatic carcinoma (PC) in c o m p a ~ n with 200 healthy controls subjects representing the Italian population. In all these patients we determined the serum level of the CA 19-9 and the Lewis phenothy~ on the erythrocytes. Results: 22 patients (39%) were of the Lewis phenotype Le(a-b-), I1 (19%) Le(a + b-), 22 (39%) Le(a-b + ) and 2 (3%) Lewis(a + b + ).The frequency of the phenothype Le(a-b-) was significantly increased (p<0.00001) compared with 200 healthy' controls. In patients affected by acute recurrent pancreatitis that phenotype was morn frequent (55%) than in patients affected by CP (28%) or by PC(30%) (Tab.).The serum levels of the CA 19-9 were not statistically diff':rent among Le phenotypes. Inside every single disease (AP, CP, and PC), we did not hnd statistlcafiy significant correlation between serum levels of CA 19-9 and the difterent phenots,~s Le. Conclusions: The frequency of phenothype Le(a-b-) is increased m a consecutive series of patients suffering fi'om pancreatic diseases compared with the general population, particularly' in ARP group. The serum levels of the CA 19-9 are not significantly different among phenotypes Le.
W1250 Oxidative Strew-Related Gene Po|ymorphisms and Susceptibility to Barrett's Esophagus Elena Piazuelo, Maria Asuncion Garcia-Gonzalez, Mark Strunk, Javier Alcedo, Juan Arenas, Angel Eerrandez, Pilaf Jimenez, Angel Lanas Both experimental and human studies have demonstrated that free radicals are involved m the ethiopathogenesis of Barretfs esophagus (BE). Antioxidant enz}anes such as superoxide dismutase and catalase plays a pivotal role in protecting esophageal mucosa from the deleterious effects of ROS. Polymorphic variants of the genes encoding these enzymes aftectnig their activity or expression may result in predisposition to Barrett's esophagus. Objective: to determine whether Ala16Val polymorphism in the manganese superoxide dismutase (MnSOD) gene and -262 T/C in the catalase gene are associated with susceptibility to Barmtt's esophagus. Methods: 107 unrelated Spanish Caucasian patients with BE and 286 ethnically matched healthy controls (HC) were included. The diagnosis of BE was established on the basis of endoscopical and histopathological criteria, Genomic DNA was extracted from peripheral blood of cases and controls. Ala 16Val MnSOD and -262T/C catalase gene polymorphisms were determined by RFLP-PCR. Clinical and epideminlogical variables were also collected by structured interview. Statistical analysis was perfbrmed by chi-sqnare test. Results: There were no differences in carriage, genotype, and allele frequencies of Ala16Val MnSOD gene polymorphism between BE patients and HC. However, the dist.qbntion of catalase genotypes was different between BE (CC: 46.2%; CT: 45.3%; TT: 8.5%) and controls (CC: 59.7%; CT: 36.8%; TT: 3.9%) p = 0.03, and carriers of the T allele were more frequent in the group of patients with BE (53.8 %) than in the control group (40.7%).; p = 0.02; R.R, = 1.69 (I.C. = 1.08-2.65). Conclusion: These data suggest that polymorphic variant 262T of catalase gene may be implicated in the susceptibility to Barrett's esophagus.
Prevalence of phenotype Le in different type of pancreatic diseases (PD) Phenotype Le
ARP
a.l~ ~.
12(55%) 6(27%) 4(16%)
a+b*
0,
CP
7(28%) 12(48%) 4(16%) 2(8%)
PC
3(30%) 4(40%} 3(30% 0
All PO
Population
22(39%) 22(39%) 11(19%)
9(4.5%) 147(73.5%) 44(22%)
2(3%) . . . .
0
Wll16 Associated Diseases With Antoimmune Pancreatitis Temmi Kamisawa, Naoto Egawa, Masakatsu Matsukawa Background/Aim: Autoimmune pancreatitis (ALP)sometimes accompanies with other autoimmune diseases or diabetes mellitus (DM). We examined associated diseases and disorders of pancreatic endocrine and exocrine tunction, and function of salivaD" glands in patients with AlP, and transition of these fl.mctions before and after steroid therapy,. Patients: Nineteen patients were diagnosed as AIP according to the following criteria; swelling of the pancreas, irregularly narrowing of the main pancreatic duct, hypergammagloburinemia or autoantibodies, and histologic findings. Steroid therapy, was ettectwe in all 9 patients treated. Methods: We examined blood glucose in hunger, 75-g oral glucose tolerance test, glycosykated hemoglobin values (HbAlc), or urinary C-peptide for pancreatic endocrine function in all patients, and periormed BT-PABA test or secretin test tbr pancreatie exocrine function in 8 patients. For salivary gland function, we measured concentrations of Na + (reflecting function of reabsorption)and beta2 microglobulin (reflecting degree of inflammatinn) in saliva and examined salivary gland scintigraphy with 99mTc in 8 patients. Results: Associated autoimmune diseases were retroperitoneal fibrosis (2 cases), rheumatoid arthritis (2), selemsing cholangitis (2), S/ogren's syndrome (1) and Sarcoidosis (1). Age of patients with these diseases (average 7 0 0 yr.) was older than 65.2 yr. of the others. Eight patients had DM. Seven patients were diagnosed as DM at the same time as AIP, and the other showed exacerbation of DM with AIP. Pancreatic exocrine dysfunction was detected in 7 of 8 cases examined After steroid therapy,, DM improved in four patients, whose pancreatic exocrine fimction also improved. Concentration of Na + and beta2 micmglobulin in saliva of patients with AIP was 22.5+/-9.1 mBoe4 (mean+/-SD) and Z65+/-1.27 mg/dl, which was significantly higher than 13.6 +/-8.4 mE@ and 1.02 +/-0.57 mg/l in a control group respectively (p<0 05). In parotid gland scintigraphy, ratio of cumulative peak count to injected radioisotope, and ratio of washout in patients with AlP was 19.9+/-11.8 % and 51.7+/-32.0 %, significantly lower than 35.3+/-84 % and 8 7 3 + / - 9 . 1 % in a control group, respectively (p<001 ) Impaired salivary gland tunction also improved after steroid therapy. Conclusions: AIP is sometimes associated with mufiifocal fibrusclerosis. Functions of pancreatic endocrine and exocrine, and sahvaD' gland are sometimes impaired, but often improve by steroid fherapy
W1251 Acid and Bile Exposure Cause Additive MAPK Activation and Proliferation in a Barrett's Adenocarcinoma Cell Line George A. Sarosi Jr,, Christie Lopez-Guzman, Stuart Spechier, Rhonda Souza Patients with Barrett's esophagus (BE) have significantly more acid and bile reflux than patients who have gastroesophageal reflux without BE. Exposure to either acid or bile salts alone has been shown to increase profiteration in explants of BE, and in a Barrett's-associated adenocarcinoma cell line (SEG-1). Recent studies suggest that acid and bile indMdually increase proliferation in BE by activating mitogen-activated protein kinases (~,~PKs) such as ERK and p38. Although it has been reported that the combination of acid and bile salts together decrease proliferation in BE, this finding seems counterintuitive. We hypothesized that bile and acid exposure together should have additive effects in activating MAPK signaling and stimulating proliferation in BE, and we tested our hypothesis in SEG-1 ceils. Methods: SEG-1 cells were grown using standard tissue culture techniques. Cells were exposed to the conjugated bile salt glyco-chenodeoxycholic acid (GCDA) at pH 7.4 (bile alone) or at pH 6.0 (bile and acid exposure) for 20 minutes to model reflux episodes. MAPK actMties were measured by immuno-blotting kinase assays. Proliferation was measured by an MTT incorporation assay, Results: Exposure of SEG-1 cells to bile alone over the physiologically relevant range of 50-500 tiM GCDA produced rapid, dose-dependent activation of both the ERK and p38 MAPKs. Exposure to bile and acid produced marked (2 to 4 fold) increases in both ERK and p38 activation beyond that seen with either acid or bile salt exposure alone. Furthermore, exposure to bile and acid produced an additive increase in cell number at 24 hours when compared to either bile salt or acid exposure alone (Table 1)(*, P< 0.05 vs 2001*M GCDA pH 7.4)Conchisions: Combined acid and bile salt exposure produces significant increases in MAPK activation and proliferation in Barmtt's adenocarcinoma cells, beyond that seen with either acid or bile salts alone. These data do not support the published contention that the combined reflux of acid and bile decreases proliferation in Barrett's esophagus.
W1249
Treatme~ Condlton Control
500pM GCDA pH 6.0 100~-2 1247 1216 1176 141 ~5' 132~7 Data expressedas cell number as a percentof ~n~I,,SEM (*,P<0,05 vs 200pM GCDA pH 7,4)
Ligand for Peroxisome Proliferator-Activated Receptor-[' Inhibits Proliferation and Induces Apoptosis of Barrett's Adenocarcinoma Both In Vitro and In Vivo Tsuyoshi Hayakawa, Yasuhiru Fujiwara, Takashi Takashima, Harunori Fujita, Masaki Hamaguchi, Masatsngn Shiba, Kazunari Tominaga, Toshio Watanabe, Nobuhide Oshitani, Kazuhide Higuchb Takayuki Matsumoto, Tetsuo Arakawa
pH 6.0 No Bile
200pM GCDA pH L4
50pM GCDA pH6.0
200pM GCDA pH 6.0
Pemxisume proliferator-activated receptor gamma (PPARy), a member of the nuclear hormone receptor SUlxrfamily, is nivolved in suppression of grmvtb of several tumors. Previous our studies showed that PPAR"/ligands inhibited growth and proliferation of Barrett's adenocarcinuma call lines (TE-7) through induction of G1 cell cycle arrest and apoptosis. The aim of this stud}, was to examine whether PPARy ligand (troglitazone) affects proliferation and apoptosis of Barmtt's adenocaminoma in vivo. Method: 1) in vitro: Human Barrett's adenocarcinoma cells (OE-33) were used. PPARy expression was determined by RT-PCR and western blotting. Efl~cts of troglitazone on proliferation and apoptosis of OE-33 were
A-633
AGA
Abstracts