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Gastric (GC) and non gastric cancer (NGC). The role of of helicobacter pylori (HP) infection
A548
AGA ABSTRACTS
GASTROENTEROLOGY, Vol. t 0 8 , No. 4
GASTRIC (GC) AND NON GASTRIC CANCER (NGC). THE ROLE OF OF HELICOBACTER PYLORI (HP) INFECTION. D Vain. M. Menegatti, M. Miglioli, M. Vergura, P. Azzarone, C. Ricci, F. Landi, 1j. Holton, G. Biasco, A. Petmnelli, B. Massardi, L. Gatta, 2R. Gusmaroli, 2F. Milesi, , 3p. Maiolo, 3A. Casadei, 4M. Rodi, 4W Giorcelli, 5A. Zambelli, 5G. Lupinaeci, F. Paunuti, L. Barbara. 1st Medical Clinic, University o3f Bologna & Oncolog,j Dept. Bologna; 2~3I Sections: Trevlglio(BO) G.B.Morgagni, Forll, *S. Andrea, Vercelli, aMaggiore, Crema, Italy; iDept. of Microbiology, University College London, UK. Aim: To assess HP semprevalence in different cancer population. Methods:During a period of 36 months 363 GC (M/F:218/145, age range 19-94; mean 69 yrS) (216 antrum, llO corpus and 37 fundus) and 305 NGC pts0ung(L)=82,breast(B)=52,genitourinary(G)=78, GI tract(GI)=63, others(O) =30) (M/F:162/143, age range 31-81; mean 57 yrs) were screened via IgG ELISA. Results: The overall seroprevalence of HP infection in GC and in NGC was 83%(303/363) and 66%(201/305) respectively (P<0.001). Table 1 shows the liP seroprevalence according to age. .............................................................................................
GC
NOC
...............................................................
HP+ve<60yrs HP+ve>6Oyrs
P " .............................
63(87%) 240(83%)
101(61%) 100(71%)
0.001 O.Ol
...............................................................................................
Table 2 shows the HP semprevalence according to cancer site. ..........................................................................................
Stomach
;
L
B
:.~.--
G
GI
O
Ti% .......
In GC no difference in HP semprevalence were found according to cancer site (83%, 82% and 81%: antrum, corpus and fundus respoctively) or histology (274 GC (233 HP+ve) were found to have an intestinal and 89 (76 HP+ve) a diffuse type(NS). Conclusion: 1. The HP seroprevalence 'in GC is statisticallyhigher compared to NGC pts and is not age related. 2. Among NGC the higher HP seroprevalence is in cancer of GI tract other than gastric and is similar to GC. 3. In GC no difference in HP seroprevalence were found according to site or histology of tumor.
DECREASED EXPRESSION OF mRNA ENCODING B14 GALACTOSYL TRANSFERASE (Gal 1") IN ADENOCARCINOMAS OF THE ESOPHAGUS. S Vasudevan ~, A Casson 2, M Khandelwal3, J Minta4, PM Rao 4, S Raja akshm 4, N Marcon s, and DSR Sarma 4. Dept of Med., Toronto Hospital'; Dep of Thoracic Surgery, Mount Sinai2; Division of Gastroenterology, The Wellesley Hospitala; Dept of Pathology, University of Toronto; Toronto, Canada. Glycosylation is needed to confer biological activity to a number of protein and lipid molecules. An alteration in this glycosylation pattern has been shown to play a pivotal role in cell to cell communication and cancer development. AI.__M:To determine the expression of Gal T, an enzyme involved in the transfer of galactosyl moiety to the nonreducing terminal of N-actylglucosamine available in the glycoproteins and glycolipids, in human esophageal carcinomas. METHODS: Endoscopic human esophageal specimens of 5 adenocarcinomas (AC), 1 squamous cell carcinoma (SCC), and surrounding normal tissue (N) were snap frozen for analysis, and a portion sent for H&E staining. Total RNA from individual specimens was purified using guanidine isothiocyanate extraction technique. Gal T transcripts were amplified by RT PCR technique and analyzed by Southern blot using radiolabelled Gal T probe. RESULTS: A significant decrease in the amounts of mRNA coding for Gal T in all AC specimens was noted compared to the surrounding N, and 1 specimen of SCC showed a similar pattern. CONCLUSION: Profound changes in the glycosylation enzymes have been reported in human hepatocellular carcinomas and rat liver with a significant decrease in the activity of Gal T during the liver cancer development. Our results show a similar pattern of glycosylation which, arising in different types of cancers, suggests that this may be a characteristic malignant phenotype. Frequent deletion of chromosome 9, which harbors Gal T, recently has been reported in esophageal AC. The decrease of Gal T reported in our results could similarly be related to a loss of chromosome 9. The pattern of expression of other glycosylation genes in esophageal Barrett's and carcinoma is under investigation.
OTONUDEX (ZD1694), A NOVEL AND S P E C I F I C T H Y M I D Y L A T E S Y N T H A S E INHIBITOR HAS P R O M I S I N G ACTIVITY IN A D V A N C E D COLORECTAL CANCER. E. Van Cutsem, D. Cunningham, 3. Zalcberg, E. Francois, J.H. Schornage], A. Adenis, M. Green, H. Starkhammer. 9. Azab. the T O M U D E X C o l o r e c t a ] C a n c e r Study Group and Z E N E C A P h a r m a c e u t i c a l s , M a e c l e s f i e ] d . U.K. TOMUDEX ZD1694 is the p r o d u c t of t a r g e t e d research programme s e e k i n g for novel p o m e n r s p e c i f i c inhibitors of thymidy]ate synthase (TS}. High s p e c i f i c i t y would abolish the indlrecr e f f e c t s on n u o l e o t i d e s y n t h e s i s seen with other drugs that i n h i b i t this enzyme. The drug e n t e r e d Phase II s t u d i e s in 1992. P r e l i m i n a r y r e s u l t s in a d v a n c e d co]orecta] c a n c e r s u g g e s t e d h i g h a c t i v i t y and T O M U D E X e n t e r e d into an e x t e n d e d P h a s e [I study fn a d v ~ c e d co]oreetal cancer in M a r c h 1993 az a dose 3f S m~/m as a short iv infusion, g i v e n S weekly. 17C patients with a d v a n c e d or m e t a s t a t l c CRC were e n t e r e d inr( the study; 6 0 % were male; mean age was 61 years (30-81 years). Over 80% ~f patients had liver m e t a s z a e e s and 30% had additional extrahepstie mezasTases All p a t l e n t s had m e a s u r a b l e ~arker disease; r e s p o n s e s were a s s e s s e d a c c o r d i n g co UICC criteria, were verified against s o u r c e data and were r e v i e w e d by an i n d e p e n d e n t panel. 26% of patlents r e s p o n d e d 9 5 % C . I . 19-33~ : in a p a r z e n L s the reszonse was complete. Minor responses were seen in an additions] patmenzs. R e s p o n s e s w e r e early, o c c u r r i n g aT first objective response assessmenz after 2 cycles of T O M U D E X in the m a j o r i t y of p a t i e n t s and o c c u r r e d in h e p a t i c and e x t r a h e p a t l c elzes. More than 8 0 % of patients were able to r e c e l v e their dose of T O M U D E X w i t h o u t s i g n i f i c a n t delay or reduction. WHO Grade S or a leueopenla, diarrhoea and nausea/vomiting each o c c u r r e d in 12% or less of patients. Clinically insignificant r e v e r s i b l e rlses in t r a n s a m i n a s e s occurred. O t h e r T o x i o i t i e s were unusual. Two toxic d e a t h s occurred (1%). both involved a combination of severe ]iarrhoea and ]eucopenla. T O M U D E X ZD1694 thus d e m o n s t r a t e s substantial a c t i v i t y in a d v a n c e d CBC and has an a c c e p t a b l e and manageable toxicity profile and a convenient a d m i n i s t r a t i o n schedule. "TOMUDEX"
is a trademark,
the p r o p e r ~ y
of ZENECA
Limited.
INVESTIGATION OF INTRACELLULAR PATHWAYS MEDIATING REGULATION OF A SIALYLTRANSFERASE GENE BY nBUTYRATE IN NORMAL AND TRANSFORMED COLONIC AND HEPATIC CELLS. R. VemulaDalli. M. Li, V. Andersen, P. Lance. GI Division, VA Medical Center and State University of New York, Buffalo, NY. n-Butyrate (nB) induces differentiation-like changes in vitro, including alterations to oligosaccharides (N-glycans) of m e m b r a n e glycoproteins and regulates expression of various genes; we reported nB-mediated inhibition of N-glycan ~2,6-sialyltransferase (ST6N) mRNA in Hep G2 cells. We report here studies in additional cell types and an investigation of potential pathways for intracellular transduction ofnB effects: METHODS: The cell system s were T84 (human colonic metastasis) and NCM 460 (a gift from Dr. M-PI Moyer, established from normal h u m a n colonocytes) cell lines and primary cultures of normal murine or rat hepatocytes, isolated by collagenase perfusion. Cells were cultured with 0,5 mM nB for up to 72 h. Levels of ST6N product were assessed by affinity chromatography with Sarnbucus nigra lectin (binds terminal = 2,6sialic acids), and of ST6N mRNA by Northern analysis. Possible mechanisms for transduction of nB effects were explored with inhibitors of protein kinase A (PKA) and protein kinase C (PKC). The role of Ca 2÷ was investigated by a fluorescent dye method and BAPTA-AM chelation for detection of intracellular fluctuations in Ca 2÷. RESULTS: ST6N mRNA expression was inhibite d p r o f o u n d l y in T84 and NCM 460 cells and primary hepatocyte cultures. Expression of = 2,6-sialyl-oligosaccharides in T84 cells was reduced by >50% after incubation (72 h) with 5 mM nB. Specific inhibitors of neither PKA nor PKC prevented nB-mediated reductions in ST6N mRNA level and this action of nB was not Ca~+-dependent. CONCLUSIONS: Downregulation of ST6N gene expression by nB, previously reported by us in Hep G2 cells, was found also in normal (NCM 460) and transformed (T84) h u m a n colonic cell lines (dividing cells) and normal murine or rat hepatocytes (non-dividing cells). The intracelinlar pathways by Which this regulatory action of nB i s transduced remain to be elucidated.
AGA ABSTRACTS
GASTROENTEROLOGY, Vol. t 0 8 , No. 4
GASTRIC (GC) AND NON GASTRIC CANCER (NGC). THE ROLE OF OF HELICOBACTER PYLORI (HP) INFECTION. D Vain. M. Menegatti, M. Miglioli, M. Vergura, P. Azzarone, C. Ricci, F. Landi, 1j. Holton, G. Biasco, A. Petmnelli, B. Massardi, L. Gatta, 2R. Gusmaroli, 2F. Milesi, , 3p. Maiolo, 3A. Casadei, 4M. Rodi, 4W Giorcelli, 5A. Zambelli, 5G. Lupinaeci, F. Paunuti, L. Barbara. 1st Medical Clinic, University o3f Bologna & Oncolog,j Dept. Bologna; 2~3I Sections: Trevlglio(BO) G.B.Morgagni, Forll, *S. Andrea, Vercelli, aMaggiore, Crema, Italy; iDept. of Microbiology, University College London, UK. Aim: To assess HP semprevalence in different cancer population. Methods:During a period of 36 months 363 GC (M/F:218/145, age range 19-94; mean 69 yrS) (216 antrum, llO corpus and 37 fundus) and 305 NGC pts0ung(L)=82,breast(B)=52,genitourinary(G)=78, GI tract(GI)=63, others(O) =30) (M/F:162/143, age range 31-81; mean 57 yrs) were screened via IgG ELISA. Results: The overall seroprevalence of HP infection in GC and in NGC was 83%(303/363) and 66%(201/305) respectively (P<0.001). Table 1 shows the liP seroprevalence according to age. .............................................................................................
GC
NOC
...............................................................
HP+ve<60yrs HP+ve>6Oyrs
P " .............................
63(87%) 240(83%)
101(61%) 100(71%)
0.001 O.Ol
...............................................................................................
Table 2 shows the HP semprevalence according to cancer site. ..........................................................................................
Stomach
;
L
B
:.~.--
G
GI
O
Ti% .......
In GC no difference in HP semprevalence were found according to cancer site (83%, 82% and 81%: antrum, corpus and fundus respoctively) or histology (274 GC (233 HP+ve) were found to have an intestinal and 89 (76 HP+ve) a diffuse type(NS). Conclusion: 1. The HP seroprevalence 'in GC is statisticallyhigher compared to NGC pts and is not age related. 2. Among NGC the higher HP seroprevalence is in cancer of GI tract other than gastric and is similar to GC. 3. In GC no difference in HP seroprevalence were found according to site or histology of tumor.
DECREASED EXPRESSION OF mRNA ENCODING B14 GALACTOSYL TRANSFERASE (Gal 1") IN ADENOCARCINOMAS OF THE ESOPHAGUS. S Vasudevan ~, A Casson 2, M Khandelwal3, J Minta4, PM Rao 4, S Raja akshm 4, N Marcon s, and DSR Sarma 4. Dept of Med., Toronto Hospital'; Dep of Thoracic Surgery, Mount Sinai2; Division of Gastroenterology, The Wellesley Hospitala; Dept of Pathology, University of Toronto; Toronto, Canada. Glycosylation is needed to confer biological activity to a number of protein and lipid molecules. An alteration in this glycosylation pattern has been shown to play a pivotal role in cell to cell communication and cancer development. AI.__M:To determine the expression of Gal T, an enzyme involved in the transfer of galactosyl moiety to the nonreducing terminal of N-actylglucosamine available in the glycoproteins and glycolipids, in human esophageal carcinomas. METHODS: Endoscopic human esophageal specimens of 5 adenocarcinomas (AC), 1 squamous cell carcinoma (SCC), and surrounding normal tissue (N) were snap frozen for analysis, and a portion sent for H&E staining. Total RNA from individual specimens was purified using guanidine isothiocyanate extraction technique. Gal T transcripts were amplified by RT PCR technique and analyzed by Southern blot using radiolabelled Gal T probe. RESULTS: A significant decrease in the amounts of mRNA coding for Gal T in all AC specimens was noted compared to the surrounding N, and 1 specimen of SCC showed a similar pattern. CONCLUSION: Profound changes in the glycosylation enzymes have been reported in human hepatocellular carcinomas and rat liver with a significant decrease in the activity of Gal T during the liver cancer development. Our results show a similar pattern of glycosylation which, arising in different types of cancers, suggests that this may be a characteristic malignant phenotype. Frequent deletion of chromosome 9, which harbors Gal T, recently has been reported in esophageal AC. The decrease of Gal T reported in our results could similarly be related to a loss of chromosome 9. The pattern of expression of other glycosylation genes in esophageal Barrett's and carcinoma is under investigation.
OTONUDEX (ZD1694), A NOVEL AND S P E C I F I C T H Y M I D Y L A T E S Y N T H A S E INHIBITOR HAS P R O M I S I N G ACTIVITY IN A D V A N C E D COLORECTAL CANCER. E. Van Cutsem, D. Cunningham, 3. Zalcberg, E. Francois, J.H. Schornage], A. Adenis, M. Green, H. Starkhammer. 9. Azab. the T O M U D E X C o l o r e c t a ] C a n c e r Study Group and Z E N E C A P h a r m a c e u t i c a l s , M a e c l e s f i e ] d . U.K. TOMUDEX ZD1694 is the p r o d u c t of t a r g e t e d research programme s e e k i n g for novel p o m e n r s p e c i f i c inhibitors of thymidy]ate synthase (TS}. High s p e c i f i c i t y would abolish the indlrecr e f f e c t s on n u o l e o t i d e s y n t h e s i s seen with other drugs that i n h i b i t this enzyme. The drug e n t e r e d Phase II s t u d i e s in 1992. P r e l i m i n a r y r e s u l t s in a d v a n c e d co]orecta] c a n c e r s u g g e s t e d h i g h a c t i v i t y and T O M U D E X e n t e r e d into an e x t e n d e d P h a s e [I study fn a d v ~ c e d co]oreetal cancer in M a r c h 1993 az a dose 3f S m~/m as a short iv infusion, g i v e n S weekly. 17C patients with a d v a n c e d or m e t a s t a t l c CRC were e n t e r e d inr( the study; 6 0 % were male; mean age was 61 years (30-81 years). Over 80% ~f patients had liver m e t a s z a e e s and 30% had additional extrahepstie mezasTases All p a t l e n t s had m e a s u r a b l e ~arker disease; r e s p o n s e s were a s s e s s e d a c c o r d i n g co UICC criteria, were verified against s o u r c e data and were r e v i e w e d by an i n d e p e n d e n t panel. 26% of patlents r e s p o n d e d 9 5 % C . I . 19-33~ : in a p a r z e n L s the reszonse was complete. Minor responses were seen in an additions] patmenzs. R e s p o n s e s w e r e early, o c c u r r i n g aT first objective response assessmenz after 2 cycles of T O M U D E X in the m a j o r i t y of p a t i e n t s and o c c u r r e d in h e p a t i c and e x t r a h e p a t l c elzes. More than 8 0 % of patients were able to r e c e l v e their dose of T O M U D E X w i t h o u t s i g n i f i c a n t delay or reduction. WHO Grade S or a leueopenla, diarrhoea and nausea/vomiting each o c c u r r e d in 12% or less of patients. Clinically insignificant r e v e r s i b l e rlses in t r a n s a m i n a s e s occurred. O t h e r T o x i o i t i e s were unusual. Two toxic d e a t h s occurred (1%). both involved a combination of severe ]iarrhoea and ]eucopenla. T O M U D E X ZD1694 thus d e m o n s t r a t e s substantial a c t i v i t y in a d v a n c e d CBC and has an a c c e p t a b l e and manageable toxicity profile and a convenient a d m i n i s t r a t i o n schedule. "TOMUDEX"
is a trademark,
the p r o p e r ~ y
of ZENECA
Limited.
INVESTIGATION OF INTRACELLULAR PATHWAYS MEDIATING REGULATION OF A SIALYLTRANSFERASE GENE BY nBUTYRATE IN NORMAL AND TRANSFORMED COLONIC AND HEPATIC CELLS. R. VemulaDalli. M. Li, V. Andersen, P. Lance. GI Division, VA Medical Center and State University of New York, Buffalo, NY. n-Butyrate (nB) induces differentiation-like changes in vitro, including alterations to oligosaccharides (N-glycans) of m e m b r a n e glycoproteins and regulates expression of various genes; we reported nB-mediated inhibition of N-glycan ~2,6-sialyltransferase (ST6N) mRNA in Hep G2 cells. We report here studies in additional cell types and an investigation of potential pathways for intracellular transduction ofnB effects: METHODS: The cell system s were T84 (human colonic metastasis) and NCM 460 (a gift from Dr. M-PI Moyer, established from normal h u m a n colonocytes) cell lines and primary cultures of normal murine or rat hepatocytes, isolated by collagenase perfusion. Cells were cultured with 0,5 mM nB for up to 72 h. Levels of ST6N product were assessed by affinity chromatography with Sarnbucus nigra lectin (binds terminal = 2,6sialic acids), and of ST6N mRNA by Northern analysis. Possible mechanisms for transduction of nB effects were explored with inhibitors of protein kinase A (PKA) and protein kinase C (PKC). The role of Ca 2÷ was investigated by a fluorescent dye method and BAPTA-AM chelation for detection of intracellular fluctuations in Ca 2÷. RESULTS: ST6N mRNA expression was inhibite d p r o f o u n d l y in T84 and NCM 460 cells and primary hepatocyte cultures. Expression of = 2,6-sialyl-oligosaccharides in T84 cells was reduced by >50% after incubation (72 h) with 5 mM nB. Specific inhibitors of neither PKA nor PKC prevented nB-mediated reductions in ST6N mRNA level and this action of nB was not Ca~+-dependent. CONCLUSIONS: Downregulation of ST6N gene expression by nB, previously reported by us in Hep G2 cells, was found also in normal (NCM 460) and transformed (T84) h u m a n colonic cell lines (dividing cells) and normal murine or rat hepatocytes (non-dividing cells). The intracelinlar pathways by Which this regulatory action of nB i s transduced remain to be elucidated.