- Email: [email protected]
zinc-superoxide dismutase levels in the gastric mucosa
Abstracts/Netherlands
Journal
oesophageal reflux, not by affecting TLESR frequency or by its effect on LES pressure, but by increasing the percentage of TLESR’s associated with pH < 4, possibly due to more acidic refluate.
of Medicine
Decreased E-cadherin expression is aswcinted with poor progaosis in patients with Barrett’s adenocarcinomas. K.K. Krishnadath, H.W. Tilanus, M. van Blankenstein, F.T. Bosman. The Oesophageal Tumour Study Group, Erasmus University,
Epithelial cell proliferative activity of Barrett’s oesopbagus: correlation with traditional cancer risk markers. F.T.M. Peters ‘, S. Ganesh ‘, E.J. Kuipers 3, A. de Jager-Krikken ‘, A. Karrenbeld ‘, W.J. Sluiter ‘, M.J. Hardonk ‘, E.C. Klinkenberg-Knol 3, C.B.H.W. Lamers 2, J.H. Kleibeuker ‘. University Hospitals, Amsterdam,
’ Groningen and ’ Leiden, Netherlands.
3 Free
University
Hospital,
The malignant potential of Barrett’s oesophagus (BE) is characterized by the presence of specialized columnar epithehum and a degree of dysplasia, both parameters being subject to bias and difficult to quantify. Medical intervention does not consistently reduce the area of BE, but may otherwise reduce its malignant potential. We evaluated epithelial cell proliferative activity of BE as a potential parameter for use in medical intervention studies. Methods: From 56 patients we obtained 109 sets of 3 biopsies of BE, each set from one level, with 3-cm intervals between sets. Cell proliferation was determined by incubation of biopsies with 5-bromo-2-deoxyuridine (BrdU) and immuno-histochemistry of BrdU-labeled nuclei, and expressed as labeling index (LI, %l. LI was determined for both surface and crypt epithelium; for each value 4000 nuclei were counted. Adjoining sections were stained with HE and PAS. Metaplasia was classified as gastric fundic (GF), gastric junctional (GJ) or specialized columnar (SC), dysplasia as absent, indefinite, low or high grade. Additional parameters included symptoms, length of BE, degree of endoscopic oesophagitis and of acid reflux. Results: LI of SC type was higher than that of GF and GJ type, the difference in the crypts being significant (see table). Median LI
GF
GJ
SC
P
Surface Crypts
0.72 2.48
1.04
1.42
NS
3.71
6.01
< 0.05
High-grade dysplasia was not present in this group. By multiple regression analysis surface LI correlated positively with degree of dysplasia (p = 0.0111, crypt LI (p = 0.000) and distance biopsy level from incisors (p = 0.041) (multiple r = 0.635). Crypt LI correlated positively with length of BE (p = 0.0331, in addition to type of metaplasia (p = 0.007) (multiple r = 0.431). Conclusion: Epithelial cell proliferative activity, as measured with BrdU-labeling, correlates well with several traditional markers of cancer risk in Barrett’s oesophagus and seems to be a useful quantitative parameter for medical intervention studies. [Supported by Dutch Cancer Society, grant GUKC 91-051.
A37
47 (I 995) Al -A42
Rotterdam,
Netherlands.
Decreased levels of the cell-cell adhesion molecule, Ecadherin, are associated with loss of differentiation in a number of carcinomas. However, the value of E-cadherin as a prognostic marker in these cancers is largely undetermined. To determine the prognostic significance of aberrant Ecadherin expression in Barrett’s adenocarcinomas, specimens were evaluated immunohistochemically for E-cadherin expression and the results were related to histological grade, tumour stage, presence of metastasis and survival. Immunohistochemistry with an E-cadherin-specific monoclonal antibody (SH91 was applied to archival tissue sections. E-cadherin expression was scored in 84 different tumour areas with varying degrees of differentiation (Gl-G3) of 39 oesophageal resection specimens containing Barrett’s adenocarcinomas. Three main patterns were found: normal E-cadherin expression ( < 10% negative cells), heterogenous expression (lo-90% negative cells) and tumour areas without E-cadherin expression (> 90% negative cells). A significant correlation was found between loss of E-cadherin expression and tumour grade (Spearman Rank corr.coef. 0.85, p < 0.0011. Also, significant correlations were found between E-cadherin expression and metastasis (Fisher exact test, p < 0.0011 and 5-year survival (log rank test, p < 0.05). Conclusions: These results suggest that E-cadherin can serve as a prognostic factor for the survival of patients with Barrett’s adenocarcinomas. Helicabacter pylori infection affects manganese- and copper/ zinc-superozide dismutase levels in the gastric mucosa. J.M. Giitz, C. van Kan, H.W. Verspaget, R.A. Veenendaal, 1. Biemond, C.B.H.W. Lamers. Department of Gastroenterology and Hepatology, Helicobacter
Unioersity Hospital, pylon’ infection is
Leiden,
Netherlands.
the most common cause of chronic gastritis. The mucosal pathology of H. pylon’ is thought to be partly due to excessive production of reactive oxygen metabolites (ROM) by phagocytes. We investigated the influence of H. pylori infection on mucosal superoxide dismutases (SOD), some major scavenger enzymes of ROM. In humans SOD is present in at least two forms, i.e. mitochondrial (Mn-SOD) and cytoplasmic (Cu/Zn-SOD). The amount and activity of both SODS were measured, respectively by ELISA and spectrophotometrical enzyme activity assay, in gastric biopsy homogenates of patients with normal mucosa (n = 391 and in patients with H. pyloti-related gastritis (n = 71). Infection and gastritis were confirmed by a combination of culture, serology and histology. The amount and activity of Mn-SOD were found to be 2to 3-fold increased (0.001 5 p I 0.051 whereas the amount and activity of Cu/Zn-SOD showed a slight decrease (p 2 0.05) in gastric mucosa of patients with H. pylon’ gastritis, in both antrum and corpus, compared to normal mucosa of patients without H. pylon’ infection. Remarkable was the observation
A38
Abstracts
/Netherlands
Journal
that the Mn-SOD level in normal biopsies of the corpus from patients with an inflamed antrum was significantly higher (p < 0.01) than that of patients with a corresponding normal antrum. Conclusions: H. pylori infection has a differential effect on mitochondrial and cytoplasmic SODS in the gastric mucosa, reflected by a marked increase in the cytokine-inducible MnSOD and a marginal decrease in the constitutive Cu/Zn-SOD. Apparently H. pylon’ infection induces an adequate response of mucosal SODS. No evidence for functional inactivation of wild-type ~53 protein by MDM-2 overexpression in gastric carcinogenesis: an immunohistocbemical study. M.E. Craanen ‘,*, P. Blok 3, W. Dekker 4, G.J.A. Offerhaus j, G.N.J. Tytgat ‘. Departments of ’ Gastroenterology and 5 Pathology, Academic Medical Centre, Amsterdam, 2 Department of Medical Oncology, The Netherlands Cancer Institute, Amsterdam, 3 Department of Pathology Westeinde Ziekenhuis, The Hague; 4 Department of Inlemai Medicine, Kennemer Gasthuis, Haarlem, Netherlands.
The p53 tumour-suppressor gene plays an important role in cell cycle control and cellular response mechanisms to DNA damage. Inactivation of wild-type p53 protein, usually not detectable by immunohistochemistry, is most commonly caused by missense and/or nonsense mutations within exons 5-8 of the p53 gene, leading to functionally inactive/altered, immunohistochemically detectable, mutant-type proteins. However, recent evidence suggests that functional inactivation of wild-type p53 protein may also result from complex formation with overexpressed MDM-2 protein as has been shown to occur in up to 30% of sarcomas. Interestingly, MDM-2 overexpression (usually resulting from MDM-2 gene amplification) was exclusively found in tumours with wild-type ~53. Since it is not known whether this latter type of p53 inactivation occurs during gastric carcinogenesis, we studied both MDM-2 overexpression and p53 protein accumulation in early gastric carcinomas (EC0 and precursor lesions. For this purpose, 45 formalin-fixed gastrectomy specimens harboring EGC were retrieved from the files. The Lauren tumour type and the presence and extent of chronic atrophic gastritis (CAG), intestinal metaplasia (IM) and dysplasia were re-assessed. IM subtypes were identified using appropriate histochemical stains. ~53 protein accumulation was assessed with DO-7 (DAKO, Denmark). A case was judged as p53positive when at least 10% of the (turnour) cells showed nuclear p53 protein accumulation. MDM-2 overexpression was assessed with a polyclonal antibody (courtesy: Dr. S. Picksley, University of Dundee, Scotland). Appropriate positive and negative controls were included for both antibodies. There were 20 intestinal-type and 25 diffuse-type EGC. p53 positivity was found in 14 (70%) intestinal-type and in 13 (52%) diffuse-type EGC. Normal gastric mucosa (n = 35) and IM foci (n = 3281, irrespective of subtype, showed complete absence of ~53 protein accumulation. In cases of dysplasia (n = 4), only severe dysplasia was p53-positive. MDM-2 overexpression was found neither in the p53-positive nor in the p53-negative lesions.
of Medicine
47 (19951 Al-A42
Conclusion: Functional inactivation of wild-type p53 protein by overexpression of MDM-2 protein does not occur during gastric carcinogenesis. Prognostic relevance of the plasminogen activation system in tissue of patients with gastric cancer. S. Ganesh, C.F.M. Sier, M.M. Heerding, G. Griffioen, C.B.H.W. Lamers, H.W. Verspaget. Department of Gastroenterology and Hepatology, University
Hospital,
Leiden,
Netherlands.
Gastric cancer has a poor prognosis and can only be cured in an early stage by surgery. Identification by pathophysiological markers of subgroups of patients with either a good or a poor prognosis might help to select patients for adjuvant therapies. Plasminogen activation (PA) parameters, important in tumour invasion and metastasis, have been shown to be of prognostic value in some human malignancies, e.g. colorectal and breast cancer. We evaluated the relation of several plasminogen activation parameters, determined by ELISAs and activity assays,in tissue homogenates of patients with gastric cancer (n = 50) with overall survival of at least 2 years, and a comparison was made with standard clinicopathological parameters. Univariate Cox analysis showed that a low tissue-type PA (t-PA) activity in normal mucosa and in carcinomas as well as a high antigen level of inhibitor type-l (PAI-1) and urokinase-type PA (u-PA) receptor in carcinomas are significantly associated with a poor overall survival. From 14 clinicopathological parameters, including age, WHO and TNM classification etc., only the number of eosinophils in the tumours was found to be associated with survival. Multivariate Cox analysis with all variables revealed that the PA parameters, apart from the u-PA receptor level in carcinoma, remained significantly (hazard-ratios: 2 I HR I 3.3, 0.02 I p I 0.05) associated with overall survival. Conclusions: Plasminogen activation parameters of both normal and carcinomatous tissue of patients with gastric cancer are of clinical relevance because of their independent prognostic impact on overall survival and might provide selection criteria for further therapy. Prevalence of p&WA cholangitis in relation bodies. M. van Milligen K. Huibregtse, G.N.J. ment dam,
of Gastroenterology, Netherlands.
in patients with primary sclerosing to other non-organ-specific autoantide Wit, J. van Bracht, E.A.J. Rauws, Tytgat, S.J.H. van Deventer. DepartAcademic
Medical
Centre,
Amster-
Primary sclerosing cholangitis (PSC) is a chronic cholestatic liver disease characterized by fibro-obliterative inflammation of the intra- and extrahepatic bile ducts. Around 70% of cases are associated with inflammatory bowel disease (IBD). As yet, the aetiology of PSC remains unknown, although the finding of cellular and humoral immune abnormalities as well as the association with certain HLA-types, all strongly point to an autoimmune-based pathogenesis. Humoral immune abnormalities in PSC patients include a high prevalence of circulating smooth muscle (SMA), anti-nuclear (ANA), and perinuclear anti-neutrophil cytoplasmic (p-ANCA) autoantibodies. We
Journal
oesophageal reflux, not by affecting TLESR frequency or by its effect on LES pressure, but by increasing the percentage of TLESR’s associated with pH < 4, possibly due to more acidic refluate.
of Medicine
Decreased E-cadherin expression is aswcinted with poor progaosis in patients with Barrett’s adenocarcinomas. K.K. Krishnadath, H.W. Tilanus, M. van Blankenstein, F.T. Bosman. The Oesophageal Tumour Study Group, Erasmus University,
Epithelial cell proliferative activity of Barrett’s oesopbagus: correlation with traditional cancer risk markers. F.T.M. Peters ‘, S. Ganesh ‘, E.J. Kuipers 3, A. de Jager-Krikken ‘, A. Karrenbeld ‘, W.J. Sluiter ‘, M.J. Hardonk ‘, E.C. Klinkenberg-Knol 3, C.B.H.W. Lamers 2, J.H. Kleibeuker ‘. University Hospitals, Amsterdam,
’ Groningen and ’ Leiden, Netherlands.
3 Free
University
Hospital,
The malignant potential of Barrett’s oesophagus (BE) is characterized by the presence of specialized columnar epithehum and a degree of dysplasia, both parameters being subject to bias and difficult to quantify. Medical intervention does not consistently reduce the area of BE, but may otherwise reduce its malignant potential. We evaluated epithelial cell proliferative activity of BE as a potential parameter for use in medical intervention studies. Methods: From 56 patients we obtained 109 sets of 3 biopsies of BE, each set from one level, with 3-cm intervals between sets. Cell proliferation was determined by incubation of biopsies with 5-bromo-2-deoxyuridine (BrdU) and immuno-histochemistry of BrdU-labeled nuclei, and expressed as labeling index (LI, %l. LI was determined for both surface and crypt epithelium; for each value 4000 nuclei were counted. Adjoining sections were stained with HE and PAS. Metaplasia was classified as gastric fundic (GF), gastric junctional (GJ) or specialized columnar (SC), dysplasia as absent, indefinite, low or high grade. Additional parameters included symptoms, length of BE, degree of endoscopic oesophagitis and of acid reflux. Results: LI of SC type was higher than that of GF and GJ type, the difference in the crypts being significant (see table). Median LI
GF
GJ
SC
P
Surface Crypts
0.72 2.48
1.04
1.42
NS
3.71
6.01
< 0.05
High-grade dysplasia was not present in this group. By multiple regression analysis surface LI correlated positively with degree of dysplasia (p = 0.0111, crypt LI (p = 0.000) and distance biopsy level from incisors (p = 0.041) (multiple r = 0.635). Crypt LI correlated positively with length of BE (p = 0.0331, in addition to type of metaplasia (p = 0.007) (multiple r = 0.431). Conclusion: Epithelial cell proliferative activity, as measured with BrdU-labeling, correlates well with several traditional markers of cancer risk in Barrett’s oesophagus and seems to be a useful quantitative parameter for medical intervention studies. [Supported by Dutch Cancer Society, grant GUKC 91-051.
A37
47 (I 995) Al -A42
Rotterdam,
Netherlands.
Decreased levels of the cell-cell adhesion molecule, Ecadherin, are associated with loss of differentiation in a number of carcinomas. However, the value of E-cadherin as a prognostic marker in these cancers is largely undetermined. To determine the prognostic significance of aberrant Ecadherin expression in Barrett’s adenocarcinomas, specimens were evaluated immunohistochemically for E-cadherin expression and the results were related to histological grade, tumour stage, presence of metastasis and survival. Immunohistochemistry with an E-cadherin-specific monoclonal antibody (SH91 was applied to archival tissue sections. E-cadherin expression was scored in 84 different tumour areas with varying degrees of differentiation (Gl-G3) of 39 oesophageal resection specimens containing Barrett’s adenocarcinomas. Three main patterns were found: normal E-cadherin expression ( < 10% negative cells), heterogenous expression (lo-90% negative cells) and tumour areas without E-cadherin expression (> 90% negative cells). A significant correlation was found between loss of E-cadherin expression and tumour grade (Spearman Rank corr.coef. 0.85, p < 0.0011. Also, significant correlations were found between E-cadherin expression and metastasis (Fisher exact test, p < 0.0011 and 5-year survival (log rank test, p < 0.05). Conclusions: These results suggest that E-cadherin can serve as a prognostic factor for the survival of patients with Barrett’s adenocarcinomas. Helicabacter pylori infection affects manganese- and copper/ zinc-superozide dismutase levels in the gastric mucosa. J.M. Giitz, C. van Kan, H.W. Verspaget, R.A. Veenendaal, 1. Biemond, C.B.H.W. Lamers. Department of Gastroenterology and Hepatology, Helicobacter
Unioersity Hospital, pylon’ infection is
Leiden,
Netherlands.
the most common cause of chronic gastritis. The mucosal pathology of H. pylon’ is thought to be partly due to excessive production of reactive oxygen metabolites (ROM) by phagocytes. We investigated the influence of H. pylori infection on mucosal superoxide dismutases (SOD), some major scavenger enzymes of ROM. In humans SOD is present in at least two forms, i.e. mitochondrial (Mn-SOD) and cytoplasmic (Cu/Zn-SOD). The amount and activity of both SODS were measured, respectively by ELISA and spectrophotometrical enzyme activity assay, in gastric biopsy homogenates of patients with normal mucosa (n = 391 and in patients with H. pyloti-related gastritis (n = 71). Infection and gastritis were confirmed by a combination of culture, serology and histology. The amount and activity of Mn-SOD were found to be 2to 3-fold increased (0.001 5 p I 0.051 whereas the amount and activity of Cu/Zn-SOD showed a slight decrease (p 2 0.05) in gastric mucosa of patients with H. pylon’ gastritis, in both antrum and corpus, compared to normal mucosa of patients without H. pylon’ infection. Remarkable was the observation
A38
Abstracts
/Netherlands
Journal
that the Mn-SOD level in normal biopsies of the corpus from patients with an inflamed antrum was significantly higher (p < 0.01) than that of patients with a corresponding normal antrum. Conclusions: H. pylori infection has a differential effect on mitochondrial and cytoplasmic SODS in the gastric mucosa, reflected by a marked increase in the cytokine-inducible MnSOD and a marginal decrease in the constitutive Cu/Zn-SOD. Apparently H. pylon’ infection induces an adequate response of mucosal SODS. No evidence for functional inactivation of wild-type ~53 protein by MDM-2 overexpression in gastric carcinogenesis: an immunohistocbemical study. M.E. Craanen ‘,*, P. Blok 3, W. Dekker 4, G.J.A. Offerhaus j, G.N.J. Tytgat ‘. Departments of ’ Gastroenterology and 5 Pathology, Academic Medical Centre, Amsterdam, 2 Department of Medical Oncology, The Netherlands Cancer Institute, Amsterdam, 3 Department of Pathology Westeinde Ziekenhuis, The Hague; 4 Department of Inlemai Medicine, Kennemer Gasthuis, Haarlem, Netherlands.
The p53 tumour-suppressor gene plays an important role in cell cycle control and cellular response mechanisms to DNA damage. Inactivation of wild-type p53 protein, usually not detectable by immunohistochemistry, is most commonly caused by missense and/or nonsense mutations within exons 5-8 of the p53 gene, leading to functionally inactive/altered, immunohistochemically detectable, mutant-type proteins. However, recent evidence suggests that functional inactivation of wild-type p53 protein may also result from complex formation with overexpressed MDM-2 protein as has been shown to occur in up to 30% of sarcomas. Interestingly, MDM-2 overexpression (usually resulting from MDM-2 gene amplification) was exclusively found in tumours with wild-type ~53. Since it is not known whether this latter type of p53 inactivation occurs during gastric carcinogenesis, we studied both MDM-2 overexpression and p53 protein accumulation in early gastric carcinomas (EC0 and precursor lesions. For this purpose, 45 formalin-fixed gastrectomy specimens harboring EGC were retrieved from the files. The Lauren tumour type and the presence and extent of chronic atrophic gastritis (CAG), intestinal metaplasia (IM) and dysplasia were re-assessed. IM subtypes were identified using appropriate histochemical stains. ~53 protein accumulation was assessed with DO-7 (DAKO, Denmark). A case was judged as p53positive when at least 10% of the (turnour) cells showed nuclear p53 protein accumulation. MDM-2 overexpression was assessed with a polyclonal antibody (courtesy: Dr. S. Picksley, University of Dundee, Scotland). Appropriate positive and negative controls were included for both antibodies. There were 20 intestinal-type and 25 diffuse-type EGC. p53 positivity was found in 14 (70%) intestinal-type and in 13 (52%) diffuse-type EGC. Normal gastric mucosa (n = 35) and IM foci (n = 3281, irrespective of subtype, showed complete absence of ~53 protein accumulation. In cases of dysplasia (n = 4), only severe dysplasia was p53-positive. MDM-2 overexpression was found neither in the p53-positive nor in the p53-negative lesions.
of Medicine
47 (19951 Al-A42
Conclusion: Functional inactivation of wild-type p53 protein by overexpression of MDM-2 protein does not occur during gastric carcinogenesis. Prognostic relevance of the plasminogen activation system in tissue of patients with gastric cancer. S. Ganesh, C.F.M. Sier, M.M. Heerding, G. Griffioen, C.B.H.W. Lamers, H.W. Verspaget. Department of Gastroenterology and Hepatology, University
Hospital,
Leiden,
Netherlands.
Gastric cancer has a poor prognosis and can only be cured in an early stage by surgery. Identification by pathophysiological markers of subgroups of patients with either a good or a poor prognosis might help to select patients for adjuvant therapies. Plasminogen activation (PA) parameters, important in tumour invasion and metastasis, have been shown to be of prognostic value in some human malignancies, e.g. colorectal and breast cancer. We evaluated the relation of several plasminogen activation parameters, determined by ELISAs and activity assays,in tissue homogenates of patients with gastric cancer (n = 50) with overall survival of at least 2 years, and a comparison was made with standard clinicopathological parameters. Univariate Cox analysis showed that a low tissue-type PA (t-PA) activity in normal mucosa and in carcinomas as well as a high antigen level of inhibitor type-l (PAI-1) and urokinase-type PA (u-PA) receptor in carcinomas are significantly associated with a poor overall survival. From 14 clinicopathological parameters, including age, WHO and TNM classification etc., only the number of eosinophils in the tumours was found to be associated with survival. Multivariate Cox analysis with all variables revealed that the PA parameters, apart from the u-PA receptor level in carcinoma, remained significantly (hazard-ratios: 2 I HR I 3.3, 0.02 I p I 0.05) associated with overall survival. Conclusions: Plasminogen activation parameters of both normal and carcinomatous tissue of patients with gastric cancer are of clinical relevance because of their independent prognostic impact on overall survival and might provide selection criteria for further therapy. Prevalence of p&WA cholangitis in relation bodies. M. van Milligen K. Huibregtse, G.N.J. ment dam,
of Gastroenterology, Netherlands.
in patients with primary sclerosing to other non-organ-specific autoantide Wit, J. van Bracht, E.A.J. Rauws, Tytgat, S.J.H. van Deventer. DepartAcademic
Medical
Centre,
Amster-
Primary sclerosing cholangitis (PSC) is a chronic cholestatic liver disease characterized by fibro-obliterative inflammation of the intra- and extrahepatic bile ducts. Around 70% of cases are associated with inflammatory bowel disease (IBD). As yet, the aetiology of PSC remains unknown, although the finding of cellular and humoral immune abnormalities as well as the association with certain HLA-types, all strongly point to an autoimmune-based pathogenesis. Humoral immune abnormalities in PSC patients include a high prevalence of circulating smooth muscle (SMA), anti-nuclear (ANA), and perinuclear anti-neutrophil cytoplasmic (p-ANCA) autoantibodies. We