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Gemcitabine – Ukrain combination affects MMP9 expression in primary pancreatic adenocarcinoma cell cultures (PPCCs)
Abstracts / Pancreatology 12 (2012) 502–597
P145. Combination cytokine, chemokine and growth factor biomarkers for diagnosis of pancreatic cancer Victoria E. Shaw 1, Claire Jenkinson 1, Brian Lane 2, Bill Greenhalf 1, 2, John P. Neoptolemos 1, 2, Eithne Costello 1, 2. 1 Liverpool Cancer Research-UK Centre, Department of Molecular and Clinical Cancer Medicine, University of Liverpool, Liverpool, UK 2 Liverpool National Institute for Health Research Biomedical Research Unit, Department of Molecular and Clinical Cancer Medicine, University of Liverpool, Liverpool, UK
Introduction: Pancreatic cancer (PDAC) diagnosis is problematic; its presentation is similar to benign disease such as obstructive jaundice and chronic pancreatitis where bilirubin levels are also increased. The clinically used PDAC biomarker, CA19-9, shows raised levels with biliary obstruction, decreasing its specificity for detecting PDAC versus benign biliary obstructive diseases. Biomarkers capable of distinguishing PDAC from benign biliary obstructive diseases are needed. Aims/objectives: To identify a panel of serum cytokines, chemokines and growth factors (CCGFs) capable of detecting PDAC from benign biliary diseases, with greater sensitivity and specificity than CA19-9 alone. Patients/methods: Luminex analysis of 27 CCGFs was performed on serum from patients with PDAC (n¼122), chronic pancreatitis (n¼45) and benign biliary obstruction (n¼26). Following normalisation and log transformation, feature finding and stepwise logistic regression was undertaken to identify the optimum combination of analytes. Results were validated in two additional independent data sets. Results: Receiver Operated Characteristic (ROC) curve analysis revealed a panel of 4 CCGFs plus CA19-9 (sensitivity¼70%, specificity¼89%) that was significantly better than CA19-9 alone (sensitivity¼77%, specificity¼63%) at distinguishing PDAC from chronic pancreatitis and benign biliary disease (AUCs 0.83 versus 0.73 respectively, p value 0.0027). Two independent sample sets confirmed this with AUCs of 0.90 versus 0.63, and 0.86 versus 0.78 respectively. Conclusion: Combinations of biomarkers can distinguish PDAC from benign pancreatic disease with better specificity than CA19-9 alone. Further work is required to refine the diagnostic equation in a larger prospective data set.
545
Results: We observed a significant reduction of MMP9 expression in both PPCCs treated with Gem-Uk combination with respect to their controls and to cells treated with Gem or Uk alone (p<0.01). Moreover, drug combination reduced significantly the number cells, and modified the structure of most nuclei with respect to untreated cells. Discussion: New approaches to reduce the metastatic behavior of PDAC are warranted, and Gem-Uk showed promising results in our preclinical studies. The new computerized approach to evaluate MMP9 staining at ICC is an ease-of-use and fast method that should be further developed both in preclinical models and for IHC analyses of PDAC tissues.
P147. A relationship between different forms of non-biliare acute pancreatitis and AB0 blood groups. A. Kozachenko, M. Zbinskaja, K. Gorbenko. Kharkiv City Clinical Emergency Hospital, Ukraine Aim: Our short epidemiological study aimed to determine whether AB0 blood could increase or decrease the risk for severe form of acute pancreatitis. Patients and methods: It was one-hospital, two years study (from 1/ 01/2010 to 31/12/2011). 108 patients with severe form and 363 patients with mild form of non-biliare AP were included. These selected cohorts were compared with control group: 4660 voluntary blood donors. Results: In the control group, the frequency of blood groups was the following: 0 – 0.3365, A – 0.359, B – 0.2195, AB – 0.085. In group with severe form of AP we have a similar distribution. In group with mild form A-blood group was nonsignificantly more cases (41.87% vs 35,9 0%) and at the same time 0-blood have nonsignificaly less cases (30.85% vs 33.65%) Conclusions: The results of this study do not support an association between risk of acute pancreatitis and the AB0 blood groups in population. But as a discussion we suggest that the effect of A blood type and 0 blood type can modified the risk of severity of non-biliare acute pancreatitis.
P148. P146. Gemcitabine – Ukrain combination affects MMP9 expression in primary pancreatic adenocarcinoma cell cultures (PPCCs) N. Funel 1, L. Botta 1, A. Alvino 1, E. Giovannetti 2, L.E. Pollina 1, V. Perrone 3, N. De Lio 3, F. Mosca 3, U. Boggi 3, D. Campani 1. 1 Department of Surgery, Division of Surgical Pathology, University of Pisa, Italy 2 VU University of Amsterdam, The Netherlands 3 Department of Oncology, Division of General and transplant Surgery, University of Pisa, Italy
Introduction: Pancreatic ductal adenocarcinoma (PDAC) is among the most lethal tumors mostly because of its invasive behavior and resistance to most chemotherapy regimens. Our previous results suggested that NSC631570 (Ukrain) modulates extracellular matrix remodeling of PDAC cell lines [Funel et al., Pancreatology 2010]. Therefore, the present study investigated the modulation of key determinants of invasive behavior such as MMP9 protein by Gem-UK, using appropriate preclinical models. Methods: Two PPCCs were seeded in multi-well chamber slides (8000 each/well) and exposed to Gem[10 nM], Uk[1 mm] and their combination. After 48-h treatment the cells were stained with the polyclonal antibody (CST-Euroclone) for MMP9. Untreated cells were used to evaluate the basal level of MMP9, and not–stained cells as negative control. Protein expression levels were evaluated with novel software for image analysis, checking both nuclei and cytoplasm staining intensity. Differences in expression values were compared by t-test/ANOVA analyses.
Microvascular density (MVD) and proliferative index (PI) in pancreatic neuroendocrine tumors (PNET) tissues: Analyses by computerized analysis (CA) N. Funel 1, E. Bertolli 1, L. Novaria 2, P. Faviana 1, C. Cappelli 2, L.E. Pollina 3, U. Boggi 4, V. Perrone 4, N. De Lio 4, C. Bartolozzi 2, F. Mosca 4, F. Basolo 1, D. Campani 1. 1
University of Pisa, Department of Surgery, Pisa, Italy University of Pisa, Division of Radiology, Pisa, Italy 3 University of Pisa, Department of Laboratory Medicine and Molecular Diagnoses, Pisa, Italy 4 University of Pisa, Division of General and Transplant Surgery, Pisa, Italy 2
Introduction: Couvelard et al demonstrated that benign tumor (ADN) showed higher MVD with respect to borderline tumor (BRD) and well differentiated carcinomas (WDC). Aim of our study was to correlate MDCT post-con- trastographic patterns of PNET with their MVD, PI and biological behaviour in ADN, BRD and WDC tissues by computerized analysis (CA). Materials and methods: We compared 22 patients, who underwent quadriphasic MDCT study in early arterial (15”), pancreatic (30”), venous (70”), delayed (180”) phases. Three different post-con- trastographic patterns were identified: pattern A (early arterial/ pancreatic enhancement and rapid wash-out, 5 cases); pattern B1 (early wash-in and no wash-out, 9 cases) and pattern B2 (enhancement only in venous or delayed phase, 8 cases). CT findings were compared with MVD in surgical pathological specimens (expressed by number of vessels/surface unit after CD34
P145. Combination cytokine, chemokine and growth factor biomarkers for diagnosis of pancreatic cancer Victoria E. Shaw 1, Claire Jenkinson 1, Brian Lane 2, Bill Greenhalf 1, 2, John P. Neoptolemos 1, 2, Eithne Costello 1, 2. 1 Liverpool Cancer Research-UK Centre, Department of Molecular and Clinical Cancer Medicine, University of Liverpool, Liverpool, UK 2 Liverpool National Institute for Health Research Biomedical Research Unit, Department of Molecular and Clinical Cancer Medicine, University of Liverpool, Liverpool, UK
Introduction: Pancreatic cancer (PDAC) diagnosis is problematic; its presentation is similar to benign disease such as obstructive jaundice and chronic pancreatitis where bilirubin levels are also increased. The clinically used PDAC biomarker, CA19-9, shows raised levels with biliary obstruction, decreasing its specificity for detecting PDAC versus benign biliary obstructive diseases. Biomarkers capable of distinguishing PDAC from benign biliary obstructive diseases are needed. Aims/objectives: To identify a panel of serum cytokines, chemokines and growth factors (CCGFs) capable of detecting PDAC from benign biliary diseases, with greater sensitivity and specificity than CA19-9 alone. Patients/methods: Luminex analysis of 27 CCGFs was performed on serum from patients with PDAC (n¼122), chronic pancreatitis (n¼45) and benign biliary obstruction (n¼26). Following normalisation and log transformation, feature finding and stepwise logistic regression was undertaken to identify the optimum combination of analytes. Results were validated in two additional independent data sets. Results: Receiver Operated Characteristic (ROC) curve analysis revealed a panel of 4 CCGFs plus CA19-9 (sensitivity¼70%, specificity¼89%) that was significantly better than CA19-9 alone (sensitivity¼77%, specificity¼63%) at distinguishing PDAC from chronic pancreatitis and benign biliary disease (AUCs 0.83 versus 0.73 respectively, p value 0.0027). Two independent sample sets confirmed this with AUCs of 0.90 versus 0.63, and 0.86 versus 0.78 respectively. Conclusion: Combinations of biomarkers can distinguish PDAC from benign pancreatic disease with better specificity than CA19-9 alone. Further work is required to refine the diagnostic equation in a larger prospective data set.
545
Results: We observed a significant reduction of MMP9 expression in both PPCCs treated with Gem-Uk combination with respect to their controls and to cells treated with Gem or Uk alone (p<0.01). Moreover, drug combination reduced significantly the number cells, and modified the structure of most nuclei with respect to untreated cells. Discussion: New approaches to reduce the metastatic behavior of PDAC are warranted, and Gem-Uk showed promising results in our preclinical studies. The new computerized approach to evaluate MMP9 staining at ICC is an ease-of-use and fast method that should be further developed both in preclinical models and for IHC analyses of PDAC tissues.
P147. A relationship between different forms of non-biliare acute pancreatitis and AB0 blood groups. A. Kozachenko, M. Zbinskaja, K. Gorbenko. Kharkiv City Clinical Emergency Hospital, Ukraine Aim: Our short epidemiological study aimed to determine whether AB0 blood could increase or decrease the risk for severe form of acute pancreatitis. Patients and methods: It was one-hospital, two years study (from 1/ 01/2010 to 31/12/2011). 108 patients with severe form and 363 patients with mild form of non-biliare AP were included. These selected cohorts were compared with control group: 4660 voluntary blood donors. Results: In the control group, the frequency of blood groups was the following: 0 – 0.3365, A – 0.359, B – 0.2195, AB – 0.085. In group with severe form of AP we have a similar distribution. In group with mild form A-blood group was nonsignificantly more cases (41.87% vs 35,9 0%) and at the same time 0-blood have nonsignificaly less cases (30.85% vs 33.65%) Conclusions: The results of this study do not support an association between risk of acute pancreatitis and the AB0 blood groups in population. But as a discussion we suggest that the effect of A blood type and 0 blood type can modified the risk of severity of non-biliare acute pancreatitis.
P148. P146. Gemcitabine – Ukrain combination affects MMP9 expression in primary pancreatic adenocarcinoma cell cultures (PPCCs) N. Funel 1, L. Botta 1, A. Alvino 1, E. Giovannetti 2, L.E. Pollina 1, V. Perrone 3, N. De Lio 3, F. Mosca 3, U. Boggi 3, D. Campani 1. 1 Department of Surgery, Division of Surgical Pathology, University of Pisa, Italy 2 VU University of Amsterdam, The Netherlands 3 Department of Oncology, Division of General and transplant Surgery, University of Pisa, Italy
Introduction: Pancreatic ductal adenocarcinoma (PDAC) is among the most lethal tumors mostly because of its invasive behavior and resistance to most chemotherapy regimens. Our previous results suggested that NSC631570 (Ukrain) modulates extracellular matrix remodeling of PDAC cell lines [Funel et al., Pancreatology 2010]. Therefore, the present study investigated the modulation of key determinants of invasive behavior such as MMP9 protein by Gem-UK, using appropriate preclinical models. Methods: Two PPCCs were seeded in multi-well chamber slides (8000 each/well) and exposed to Gem[10 nM], Uk[1 mm] and their combination. After 48-h treatment the cells were stained with the polyclonal antibody (CST-Euroclone) for MMP9. Untreated cells were used to evaluate the basal level of MMP9, and not–stained cells as negative control. Protein expression levels were evaluated with novel software for image analysis, checking both nuclei and cytoplasm staining intensity. Differences in expression values were compared by t-test/ANOVA analyses.
Microvascular density (MVD) and proliferative index (PI) in pancreatic neuroendocrine tumors (PNET) tissues: Analyses by computerized analysis (CA) N. Funel 1, E. Bertolli 1, L. Novaria 2, P. Faviana 1, C. Cappelli 2, L.E. Pollina 3, U. Boggi 4, V. Perrone 4, N. De Lio 4, C. Bartolozzi 2, F. Mosca 4, F. Basolo 1, D. Campani 1. 1
University of Pisa, Department of Surgery, Pisa, Italy University of Pisa, Division of Radiology, Pisa, Italy 3 University of Pisa, Department of Laboratory Medicine and Molecular Diagnoses, Pisa, Italy 4 University of Pisa, Division of General and Transplant Surgery, Pisa, Italy 2
Introduction: Couvelard et al demonstrated that benign tumor (ADN) showed higher MVD with respect to borderline tumor (BRD) and well differentiated carcinomas (WDC). Aim of our study was to correlate MDCT post-con- trastographic patterns of PNET with their MVD, PI and biological behaviour in ADN, BRD and WDC tissues by computerized analysis (CA). Materials and methods: We compared 22 patients, who underwent quadriphasic MDCT study in early arterial (15”), pancreatic (30”), venous (70”), delayed (180”) phases. Three different post-con- trastographic patterns were identified: pattern A (early arterial/ pancreatic enhancement and rapid wash-out, 5 cases); pattern B1 (early wash-in and no wash-out, 9 cases) and pattern B2 (enhancement only in venous or delayed phase, 8 cases). CT findings were compared with MVD in surgical pathological specimens (expressed by number of vessels/surface unit after CD34