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P.08.5 IDENTIFICATION OF CDH1 GERMLINE MUTATIONS IN SPORADIC GASTRIC CANCER PATIENTS AND SUBJECTS AT RISK TO DEVELOP GASTRIC CANCER
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Abstracts of the 18th National Congress of Digestive Diseases / Digestive and Liver Disease 44S (2012) S55–S220
P.08.5 IDENTIFICATION OF CDH1 GERMLINE MUTATIONS IN SPORADIC GASTRIC CANCER PATIENTS AND SUBJECTS AT RISK TO DEVELOP GASTRIC CANCER V. De Re ∗ ,1 , M. Garziera 1 , S. Geremia 2 , L. Caggiari 1 , M. De Zorzi 1 , V. Canzonieri 1 , G. Toffoli 1 , S. Maiero 1 , R. Cannizzaro 1 1 Centro
di Riferimento Oncologico Irccs, Aviano (Pn), Italy; 2 Ceb-Centre of Excellence In Biocrystallography, Trieste, Italy Background and aim: Germline mutations in the E-cadherin (CDH1) gene are involved in the tumorigenesis of hereditary diffuse gastric cancer, but the role and the frequency of these variants in sporadic GC patients and in subjects at risk to develop GC is poorly investigated. Material and methods: 48 consecutive GC patients median age 64, 59 first degree relatives (FDR) median age 49, 20 patients affected by autoimmune atrophic gastritis (AAG) median age 56 and 52 healthy donors (HD), were enrolled for this study. Screening of the entire coding region and all exon flanking sequences of the CDH1 gene was performed by DNA direct sequencing. The potential deleterious nature of germline missense variants was explored by means of structural modelling studies and bioinformatic tools. Immunohistochemistry and characterization of functional impact on splicing were assessed. Results: All the missense mutations were found in the extracellular domain of E-cadherin protein. One novel missense mutation in exon 6 (p.G274S) with a controversial pathogenic role was found in a GC patient. Other two missense mutations already reported in GC families (p.A298T, and p.T470I), and one variant in the promoter region (+54 G>C) in two distinct patients, were found in GC group. In FDR, one mutation in the promoter region (+71 C>G), one in the untraslated region (5’UTR -176 C>T from ATG), and an intronic variant close to exon 5 described in early-onset GC (IVS4 -18 C>T). In AAG, one probable new intronic variant near to exon 1 (IVS1+7 C>T). The intronic variant close to exon 13 (IVS12 -13 T>C) was found in a GC patient and also in one control. The missense variant p.A592T, different silent variants and polymorphisms were also detected in all groups. Conclusions: Our results show that CDH1 germline variants with a potential pathological role can also be detected in sporadic GC patients without fulfilling the strict criteria for familiar GC. This data suggest the need to improve the case study to better investigate contribution of E-cadherin variants in sporadic GC patients and in general population. Moreover, our results highlight that screening for CDH1 mutations could be a tool to define a risk group of patients.
P.08.6 UROTENSIN-II RECEPTOR: A NEW MARKER FOR THE DIFFERENTIATION AND THE TARGET IN COLON CANCER? A. Federico ∗ ,1 , M. Romano 1 , A.G. Gravina 1 , M. Di Domenico 2 , M. Marra 3 , S. Zappavigna 3 , F.P. D’Armiento 4 , R. Ippolito 4 , C. Tuccillo 1 , P. Grieco 5 , E. Novellino 5 , C. Loguercio 5 , M. Caraglia 3 1 Department of Clinical and Experimental Medicine and Surgery “F Magrassi e A Lanzara”, Second University of Naples, Naples, Italy; 2 Department of General Pathology, Second University of Naples, Naples, Italy; 3 Department of Biochemistry and Biophysics, Second University of Naples, Naples, Italy; 4 Department of Biomorphological and Functional Sciences, University Federico II, Naples, Italy; 5 Department of Pharmaceutical and Toxicological Chemistry, University of Naples Federico II, Naples, Italy
Background and aim: Urotensin II (U-II) is a cyclic peptide that we have previously hypothesized to be involved in the regulation of cancer processes. U-II induces proliferative effects on human corticosurrenal cancer and this effect is mediated through the interaction with U-II receptor, a GcRP protein (UTR). UTR has been shown to be expressed in vivo in human prostate cancer tissues and it is moderately expressed in prostate hyperplasia and at high levels in well differentiated prostate adenocarcinoma. Whether UTR is expressed in colon cancer cells is unknown. Aim of this study is to evaluate: 1) UTR protein expression in a number
of human epithelial colon cancer cell lines; 2) UTR mRNA and protein expression in normal colon tissue, in adenomatous polyps and in colon cancer; 3) whether UTR agonists stimulate colon cancer cell growth. Material and methods: 1) Cell lines: LOVO, HT-29, SW620, COLO and WIDR; 2) UTR mRNA expression by RT-PCR; 3) UTR protein expression by western blot in colon cancer cells and by immunohistochemistry in normal colon tissue (7 subjects), in colonic adenomatous polyps (7 subjects), in colon carcinoma (24 subjects) and in normal colon from patients with colon cancer (24 subjects) 4) Cell proliferation by MTT assay. Results: 1) UTR protein was highly expressed in WIDR cell line whereas its expression was low in LOVO, HT-29, SW620 and COLO cells; 2) UTR mRNA expression was increased by 3 folds in patients with adenomatous polyps and by 8 folds in patients with colon cancer compared with normal (control) colon; 3) at immunohistochemistry, UTR was expressed in 5%-30% of normal colon, 30%-48% of adenomatous polyps, and in 65%-90% of colon adenocarcinomas; 4) no differences in mRNA or protein expression was found between colon from control subjects and normal colon from cancer patients; 5) agonistic peptides and U-II induced an about 20%-40% growth stimulus at a concentration close to the Kd of the receptor (about 10 nM). Conclusions: 1) UTR is expressed in colon cancer cell lines; 2) UTR is over-expressed both at the mRNA and protein level in adenomatous polyps and at a higher degree in colon adenocarcinoma; 3) UTR agonistic peptides stimulate colon cancer cell growth; 4) we hypothesize that U-II/UTR mediated pathway may play a role in colon carcinogenesis.
P.08.7 MAJOR BLEEDING EPISODES AMONG PATIENTS WITH HCC DURING SORAFENIB TREATMENT: THE DARK SIDE OF A RELATIVELY SAFE DRUG? A. De Santis, C. Iegri ∗ , S. Trapani, M. Lupo, F. Giubilo, C. Bassanelli, F. Attili, G. Gallusi, A.F. Attili Policlinico Umberto 1 - Uoc di Gastroenterologia ed Endoscopia Digestiva, Roma, Italy Background and aim: Major bleeding episodes have been described during anti-angiogenic treatment, including Sorafenib. The incidence of bleeding events (all grades) was 16.7% (95% CI 12.7–21.5) in pts with various cancers treated with tyrosine-kinase inhibitors. SHARP trial did not identify an overall increase in the risk of hemorrage and the rate of variceal bleeding was not significantly different in pts with HepatoCellular Carcinoma (HCC) during treatment with Sorafenib (2%) vs placebo group (4%).However, in SHARP study both the Sorafenib and placebo cohorts were extremely selected (95% Child Pugh A); the prevalence of oesophageal varices (OV), gastroesophageal varices GOV) or gastropathy (GP)in these patients were thus presumably low. The bleeding rate could be relatively higher in the “real clinical practice”. Our aim was to evaluate the bleeding rate in a cohorts of patients with HCC treated with Sorafenib. Material and methods: We prospectively followed up 49 patients treated with Sorafenib between August 08 and October 2011. 37 pts (76%) were male. The mean age was: 68.7 + 10.44 y. 69.4% and 30.6% of pts were CP A and B7, respectively. 57.1% and 42.9% of pts were BCLC C and B stages respectively. At baseline OV, GP and GOV were found in 19 pts (38.7%),18 pts (36.7%) and 5 pts (10.2%), respectively. Results: 12 pts (24.5%) experienced at least one GI bleeding episode during treatment.An upper gastrointestinal bleeding (UGB) occurred in 10 pts:varices
Abstracts of the 18th National Congress of Digestive Diseases / Digestive and Liver Disease 44S (2012) S55–S220
P.08.5 IDENTIFICATION OF CDH1 GERMLINE MUTATIONS IN SPORADIC GASTRIC CANCER PATIENTS AND SUBJECTS AT RISK TO DEVELOP GASTRIC CANCER V. De Re ∗ ,1 , M. Garziera 1 , S. Geremia 2 , L. Caggiari 1 , M. De Zorzi 1 , V. Canzonieri 1 , G. Toffoli 1 , S. Maiero 1 , R. Cannizzaro 1 1 Centro
di Riferimento Oncologico Irccs, Aviano (Pn), Italy; 2 Ceb-Centre of Excellence In Biocrystallography, Trieste, Italy Background and aim: Germline mutations in the E-cadherin (CDH1) gene are involved in the tumorigenesis of hereditary diffuse gastric cancer, but the role and the frequency of these variants in sporadic GC patients and in subjects at risk to develop GC is poorly investigated. Material and methods: 48 consecutive GC patients median age 64, 59 first degree relatives (FDR) median age 49, 20 patients affected by autoimmune atrophic gastritis (AAG) median age 56 and 52 healthy donors (HD), were enrolled for this study. Screening of the entire coding region and all exon flanking sequences of the CDH1 gene was performed by DNA direct sequencing. The potential deleterious nature of germline missense variants was explored by means of structural modelling studies and bioinformatic tools. Immunohistochemistry and characterization of functional impact on splicing were assessed. Results: All the missense mutations were found in the extracellular domain of E-cadherin protein. One novel missense mutation in exon 6 (p.G274S) with a controversial pathogenic role was found in a GC patient. Other two missense mutations already reported in GC families (p.A298T, and p.T470I), and one variant in the promoter region (+54 G>C) in two distinct patients, were found in GC group. In FDR, one mutation in the promoter region (+71 C>G), one in the untraslated region (5’UTR -176 C>T from ATG), and an intronic variant close to exon 5 described in early-onset GC (IVS4 -18 C>T). In AAG, one probable new intronic variant near to exon 1 (IVS1+7 C>T). The intronic variant close to exon 13 (IVS12 -13 T>C) was found in a GC patient and also in one control. The missense variant p.A592T, different silent variants and polymorphisms were also detected in all groups. Conclusions: Our results show that CDH1 germline variants with a potential pathological role can also be detected in sporadic GC patients without fulfilling the strict criteria for familiar GC. This data suggest the need to improve the case study to better investigate contribution of E-cadherin variants in sporadic GC patients and in general population. Moreover, our results highlight that screening for CDH1 mutations could be a tool to define a risk group of patients.
P.08.6 UROTENSIN-II RECEPTOR: A NEW MARKER FOR THE DIFFERENTIATION AND THE TARGET IN COLON CANCER? A. Federico ∗ ,1 , M. Romano 1 , A.G. Gravina 1 , M. Di Domenico 2 , M. Marra 3 , S. Zappavigna 3 , F.P. D’Armiento 4 , R. Ippolito 4 , C. Tuccillo 1 , P. Grieco 5 , E. Novellino 5 , C. Loguercio 5 , M. Caraglia 3 1 Department of Clinical and Experimental Medicine and Surgery “F Magrassi e A Lanzara”, Second University of Naples, Naples, Italy; 2 Department of General Pathology, Second University of Naples, Naples, Italy; 3 Department of Biochemistry and Biophysics, Second University of Naples, Naples, Italy; 4 Department of Biomorphological and Functional Sciences, University Federico II, Naples, Italy; 5 Department of Pharmaceutical and Toxicological Chemistry, University of Naples Federico II, Naples, Italy
Background and aim: Urotensin II (U-II) is a cyclic peptide that we have previously hypothesized to be involved in the regulation of cancer processes. U-II induces proliferative effects on human corticosurrenal cancer and this effect is mediated through the interaction with U-II receptor, a GcRP protein (UTR). UTR has been shown to be expressed in vivo in human prostate cancer tissues and it is moderately expressed in prostate hyperplasia and at high levels in well differentiated prostate adenocarcinoma. Whether UTR is expressed in colon cancer cells is unknown. Aim of this study is to evaluate: 1) UTR protein expression in a number
of human epithelial colon cancer cell lines; 2) UTR mRNA and protein expression in normal colon tissue, in adenomatous polyps and in colon cancer; 3) whether UTR agonists stimulate colon cancer cell growth. Material and methods: 1) Cell lines: LOVO, HT-29, SW620, COLO and WIDR; 2) UTR mRNA expression by RT-PCR; 3) UTR protein expression by western blot in colon cancer cells and by immunohistochemistry in normal colon tissue (7 subjects), in colonic adenomatous polyps (7 subjects), in colon carcinoma (24 subjects) and in normal colon from patients with colon cancer (24 subjects) 4) Cell proliferation by MTT assay. Results: 1) UTR protein was highly expressed in WIDR cell line whereas its expression was low in LOVO, HT-29, SW620 and COLO cells; 2) UTR mRNA expression was increased by 3 folds in patients with adenomatous polyps and by 8 folds in patients with colon cancer compared with normal (control) colon; 3) at immunohistochemistry, UTR was expressed in 5%-30% of normal colon, 30%-48% of adenomatous polyps, and in 65%-90% of colon adenocarcinomas; 4) no differences in mRNA or protein expression was found between colon from control subjects and normal colon from cancer patients; 5) agonistic peptides and U-II induced an about 20%-40% growth stimulus at a concentration close to the Kd of the receptor (about 10 nM). Conclusions: 1) UTR is expressed in colon cancer cell lines; 2) UTR is over-expressed both at the mRNA and protein level in adenomatous polyps and at a higher degree in colon adenocarcinoma; 3) UTR agonistic peptides stimulate colon cancer cell growth; 4) we hypothesize that U-II/UTR mediated pathway may play a role in colon carcinogenesis.
P.08.7 MAJOR BLEEDING EPISODES AMONG PATIENTS WITH HCC DURING SORAFENIB TREATMENT: THE DARK SIDE OF A RELATIVELY SAFE DRUG? A. De Santis, C. Iegri ∗ , S. Trapani, M. Lupo, F. Giubilo, C. Bassanelli, F. Attili, G. Gallusi, A.F. Attili Policlinico Umberto 1 - Uoc di Gastroenterologia ed Endoscopia Digestiva, Roma, Italy Background and aim: Major bleeding episodes have been described during anti-angiogenic treatment, including Sorafenib. The incidence of bleeding events (all grades) was 16.7% (95% CI 12.7–21.5) in pts with various cancers treated with tyrosine-kinase inhibitors. SHARP trial did not identify an overall increase in the risk of hemorrage and the rate of variceal bleeding was not significantly different in pts with HepatoCellular Carcinoma (HCC) during treatment with Sorafenib (2%) vs placebo group (4%).However, in SHARP study both the Sorafenib and placebo cohorts were extremely selected (95% Child Pugh A); the prevalence of oesophageal varices (OV), gastroesophageal varices GOV) or gastropathy (GP)in these patients were thus presumably low. The bleeding rate could be relatively higher in the “real clinical practice”. Our aim was to evaluate the bleeding rate in a cohorts of patients with HCC treated with Sorafenib. Material and methods: We prospectively followed up 49 patients treated with Sorafenib between August 08 and October 2011. 37 pts (76%) were male. The mean age was: 68.7 + 10.44 y. 69.4% and 30.6% of pts were CP A and B7, respectively. 57.1% and 42.9% of pts were BCLC C and B stages respectively. At baseline OV, GP and GOV were found in 19 pts (38.7%),18 pts (36.7%) and 5 pts (10.2%), respectively. Results: 12 pts (24.5%) experienced at least one GI bleeding episode during treatment.An upper gastrointestinal bleeding (UGB) occurred in 10 pts:varices