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Internal and external factors initiating carcinogenesis in Wistar rats
Abstracts / Pancreatology 16 (2016) S1eS130
Abstract ID: 1511. Acute pancreatitis promotes liver metastasis in a mouse model of pancreatic ductal adenocarcinoma via SDF-1_CXCR4 pathway Yonghua Chen 1, Nengwen Ke 1, Chunlu Tan 1, Wei Huang 2, Xubao Liu 1, Robert Sutton 3 1 Department of Pancreatic Surgery, West China Hospital, Sichuan University, Chengdu, China 2 Department of Integrated Traditional Chinese and Western Medicine, West China Hospital, Sichuan University, Chengdu, China 3 NIHR Liverpool Pancreas Biomedical Research Unit, Royal Liverpool University Hospital, University of Liverpool, Liverpool, United Kingdom
Introduction: Whether acute pancreatitis (AP) promotes pancreatic ductal adenocarcinoma (PDAC) liver metastasis is unknown. Aims: To investigate the role of AP in PDAC liver metastasis. Materials & methods: The effect of stromal cell-derived factor-1 (SDF1, 100 ng/ml) and peritoneal lavage fluid (PLF after PBS or caerulein 50 mg/ kg/h 8 injections) on Panc02 (PDAC cell line) migration was assessed with or without SDF-1 receptor (CXCR4) antagonist AMD3100 (500 ng/ml). Mice received intrasplenic Panc02 cells 48 h after PBS, caerulein, or caerulein and AMD3100 (5 mg/kg) injection and were sacrificed at 24 h and 48 h to identify CM-Dil labelled Panc02 cells in the liver by confocal microscopy. Further mice underwent splenectomy 48 h after Panc02 cell injection and sacrifice 3 weeks later for liver histopathology. Results: SDF-1 levels in PLF were significantly increased at 24 h and 72 h after caerulein (both P<0.01). Both SDF-1 and PLF induced significant migration of Panc02 cells vs PBS (both P<0.001), markedly inhibited by AMD3100 (both P<0.001). PBS-injected mice had few CM-Dil fluorescent cells in the liver visualised at 24 h (2.2 cells/HPF) and no increase at 48 h. Caerulein significantly induced uptake of CM-Dil fluorescent cells in the liver at 24 h (15.3 cell/HPF) and 48 h (20.4 cells/HPF, both P<0.001), but AMD3100 significantly reduced these changes (10.91/HPF and 6.8/HPF, respectively, P<0.001); histopathology at 3 weeks confirmed these findings. Conclusion: AP accelerated PDAC cell liver metastasis through a SDF1eCXCR4 mediated pathway. These data suggest AP should be avoided in PDAC.
Abstract ID: 1514. Targeting individuals with new-onset diabetes for early detection of pancreatic cancer in this high risk group
S19
pancreatitis (with/without diabetes), new-onset and long-term 2 diabetes, and healthy controls. Results: A discovery program revealed 141 markers for early detection of PDAC. IPA found an association with metabolic disease pathways in 45 of the 141 possible candidate proteins identified. Twenty-five of these were significantly enriched for an association with diabetes. Serum levels of several candidate biomarkers significantly differed in pancreatic cancerassociated diabetes (PDAC-DM) compared with diabetes (DM), further comparison of markers revealed a difference between the protein profile of PDAC and PDAC-DM patients to that of individuals with type 2 diabetes. Conclusion: Proteins associated with metabolic disease may behave differently in type 2 and type 3c diabetes, a comprehensive analysis will be necessary to identify a type 3c specific protein profile.
Abstract ID: 1524. Internal and external factors initiating carcinogenesis in Wistar rats Irina Trubitsyna 1, Anna Smirnova 1, Ludmila Vinokurova 1, Dmitry Bordin 1, Andrey Guliaev 2, Elena Dubtsova 1 1 2
Moscow Clinical Scientific Center, Russia Institute of Gene Biology, Russia
Introduction: In modelling of formation of neoplastic polyps in colon a model with 1,2-azoxymethane (AOM) is used, which initiates growth of polyps and tumors in LI, therefore affecting wnt/akt2 signalling system. The model is used to study an impact of feeding, gut microbiome etc. on growth and proliferation of tumors. Aims: We investigated an impact of AOM in combination with inflammation on development of polyps and tumors in LI. Materials & methods: We used special rat feed made by “Laboratorkorm”. Healthy female Wistar rats (IBCh RAS, Pushchino branch, n¼20, body weight 250-260 g) were divided into 4 groups: (1) intact, (2) impacted by AOM (15 mg/kg, intraperitoneally injected twice, with 14 days interval), (3) with colon ulcer (caused by acetic acid), (4) with colon ulcer and impacted by AOM. Histomorphometric studies were performed on day 70. Results: Presence of polyps and adenocarcinomas in colon as well as renal cysts and structure disturbance of pancreas were detected in groups 2 and 4. We revealed a significant increase of food consumption in groups 2 and 4. Were found cysts of pancreas in 3 animals from each group as well as visceral adipose tissue enlargement, liver fatty degeneration and growth of neoplastic polyps in colon. Conclusion: This model probably will help us to design studies of polycystic pancreas disease; however it should be improved considering influence of external factors.
Lucy Oldfield 1, Claire Jenkinson 1, Tejpal Purewal 2, Robert Sutton 1, John P. Neoptolemos 1, William Greenhalf 1, Eithne Costello 1 1 Department of Molecular and Clinical Cancer Medicine, University of Liverpool, United Kingdom 2 Department of Diabetes and Endocrinology, Royal Liverpool University Hospital, United Kingdom
Introduction: Cancer-related diabetes can precede pancreatic cancer diagnosis by up to 3 years. Only 1% of patients with new-onset diabetes have PDAC, differentiating pancreatic cancer-associated diabetes (type 3c) from the more common type 2 is essential for screening to be viable in this high risk group. Aims: To investigate whether individuals with undetected pancreatic cancer, newly diagnosed with diabetes, can be distinguished from individuals with more common new-onset type 2 disease. Materials & methods: Discovery work was performed using Luminex and isobaric tags for relative and absolute quantification (iTRAQ) on 161 pre-diagnosis and PDAC samples. Ingenuity pathway analysis (IPA) on proteins from discovery work identified those associated with metabolic disease pathways and diabetes. Candidate biomarkers were assessed using immunoblotting and/or ELISA on 135 serum samples categorised into various disease groups: PDAC (with/without diabetes), chronic
Abstract ID: 1526. Severity histologic score of acute pancreatitis in murine models: Preliminary results o Carlos Pedro Silva Vaz 1, Rui Oliveira 2, Ana Margarida Abrantes 3, Joa o 3, Anto nio Gouveia 4, Miguel Castelo-Branco 5, Maria Encarnaça o 6 e Guilherme Tralha Filomena Botelho 3, Jos 1 General Surgery Department, Unidade Local de Saúde de Castelo Branco j CICS, Portugal 2 Pathology Department, Centro Hospitalar e Universit ario de Coimbra, Portugal 3 IBILI e Institute for Biomedical Imaging and Life Sciences, Faculty of Medicine, University of Coimbra j CIMAGO, Portugal 4 General Surgery Department, Unidade Local de Saúde de Castelo Branco, Portugal 5 CICS e Health Sciences Research Centre, Faculty of Health Sciences, University of Beira Interior, Portugal 6 rio de General Surgery Department, Centro Hospitalar e Universita Coimbra j IBILI j CIMAGO, Portugal
Abstract ID: 1511. Acute pancreatitis promotes liver metastasis in a mouse model of pancreatic ductal adenocarcinoma via SDF-1_CXCR4 pathway Yonghua Chen 1, Nengwen Ke 1, Chunlu Tan 1, Wei Huang 2, Xubao Liu 1, Robert Sutton 3 1 Department of Pancreatic Surgery, West China Hospital, Sichuan University, Chengdu, China 2 Department of Integrated Traditional Chinese and Western Medicine, West China Hospital, Sichuan University, Chengdu, China 3 NIHR Liverpool Pancreas Biomedical Research Unit, Royal Liverpool University Hospital, University of Liverpool, Liverpool, United Kingdom
Introduction: Whether acute pancreatitis (AP) promotes pancreatic ductal adenocarcinoma (PDAC) liver metastasis is unknown. Aims: To investigate the role of AP in PDAC liver metastasis. Materials & methods: The effect of stromal cell-derived factor-1 (SDF1, 100 ng/ml) and peritoneal lavage fluid (PLF after PBS or caerulein 50 mg/ kg/h 8 injections) on Panc02 (PDAC cell line) migration was assessed with or without SDF-1 receptor (CXCR4) antagonist AMD3100 (500 ng/ml). Mice received intrasplenic Panc02 cells 48 h after PBS, caerulein, or caerulein and AMD3100 (5 mg/kg) injection and were sacrificed at 24 h and 48 h to identify CM-Dil labelled Panc02 cells in the liver by confocal microscopy. Further mice underwent splenectomy 48 h after Panc02 cell injection and sacrifice 3 weeks later for liver histopathology. Results: SDF-1 levels in PLF were significantly increased at 24 h and 72 h after caerulein (both P<0.01). Both SDF-1 and PLF induced significant migration of Panc02 cells vs PBS (both P<0.001), markedly inhibited by AMD3100 (both P<0.001). PBS-injected mice had few CM-Dil fluorescent cells in the liver visualised at 24 h (2.2 cells/HPF) and no increase at 48 h. Caerulein significantly induced uptake of CM-Dil fluorescent cells in the liver at 24 h (15.3 cell/HPF) and 48 h (20.4 cells/HPF, both P<0.001), but AMD3100 significantly reduced these changes (10.91/HPF and 6.8/HPF, respectively, P<0.001); histopathology at 3 weeks confirmed these findings. Conclusion: AP accelerated PDAC cell liver metastasis through a SDF1eCXCR4 mediated pathway. These data suggest AP should be avoided in PDAC.
Abstract ID: 1514. Targeting individuals with new-onset diabetes for early detection of pancreatic cancer in this high risk group
S19
pancreatitis (with/without diabetes), new-onset and long-term 2 diabetes, and healthy controls. Results: A discovery program revealed 141 markers for early detection of PDAC. IPA found an association with metabolic disease pathways in 45 of the 141 possible candidate proteins identified. Twenty-five of these were significantly enriched for an association with diabetes. Serum levels of several candidate biomarkers significantly differed in pancreatic cancerassociated diabetes (PDAC-DM) compared with diabetes (DM), further comparison of markers revealed a difference between the protein profile of PDAC and PDAC-DM patients to that of individuals with type 2 diabetes. Conclusion: Proteins associated with metabolic disease may behave differently in type 2 and type 3c diabetes, a comprehensive analysis will be necessary to identify a type 3c specific protein profile.
Abstract ID: 1524. Internal and external factors initiating carcinogenesis in Wistar rats Irina Trubitsyna 1, Anna Smirnova 1, Ludmila Vinokurova 1, Dmitry Bordin 1, Andrey Guliaev 2, Elena Dubtsova 1 1 2
Moscow Clinical Scientific Center, Russia Institute of Gene Biology, Russia
Introduction: In modelling of formation of neoplastic polyps in colon a model with 1,2-azoxymethane (AOM) is used, which initiates growth of polyps and tumors in LI, therefore affecting wnt/akt2 signalling system. The model is used to study an impact of feeding, gut microbiome etc. on growth and proliferation of tumors. Aims: We investigated an impact of AOM in combination with inflammation on development of polyps and tumors in LI. Materials & methods: We used special rat feed made by “Laboratorkorm”. Healthy female Wistar rats (IBCh RAS, Pushchino branch, n¼20, body weight 250-260 g) were divided into 4 groups: (1) intact, (2) impacted by AOM (15 mg/kg, intraperitoneally injected twice, with 14 days interval), (3) with colon ulcer (caused by acetic acid), (4) with colon ulcer and impacted by AOM. Histomorphometric studies were performed on day 70. Results: Presence of polyps and adenocarcinomas in colon as well as renal cysts and structure disturbance of pancreas were detected in groups 2 and 4. We revealed a significant increase of food consumption in groups 2 and 4. Were found cysts of pancreas in 3 animals from each group as well as visceral adipose tissue enlargement, liver fatty degeneration and growth of neoplastic polyps in colon. Conclusion: This model probably will help us to design studies of polycystic pancreas disease; however it should be improved considering influence of external factors.
Lucy Oldfield 1, Claire Jenkinson 1, Tejpal Purewal 2, Robert Sutton 1, John P. Neoptolemos 1, William Greenhalf 1, Eithne Costello 1 1 Department of Molecular and Clinical Cancer Medicine, University of Liverpool, United Kingdom 2 Department of Diabetes and Endocrinology, Royal Liverpool University Hospital, United Kingdom
Introduction: Cancer-related diabetes can precede pancreatic cancer diagnosis by up to 3 years. Only 1% of patients with new-onset diabetes have PDAC, differentiating pancreatic cancer-associated diabetes (type 3c) from the more common type 2 is essential for screening to be viable in this high risk group. Aims: To investigate whether individuals with undetected pancreatic cancer, newly diagnosed with diabetes, can be distinguished from individuals with more common new-onset type 2 disease. Materials & methods: Discovery work was performed using Luminex and isobaric tags for relative and absolute quantification (iTRAQ) on 161 pre-diagnosis and PDAC samples. Ingenuity pathway analysis (IPA) on proteins from discovery work identified those associated with metabolic disease pathways and diabetes. Candidate biomarkers were assessed using immunoblotting and/or ELISA on 135 serum samples categorised into various disease groups: PDAC (with/without diabetes), chronic
Abstract ID: 1526. Severity histologic score of acute pancreatitis in murine models: Preliminary results o Carlos Pedro Silva Vaz 1, Rui Oliveira 2, Ana Margarida Abrantes 3, Joa o 3, Anto nio Gouveia 4, Miguel Castelo-Branco 5, Maria Encarnaça o 6 e Guilherme Tralha Filomena Botelho 3, Jos 1 General Surgery Department, Unidade Local de Saúde de Castelo Branco j CICS, Portugal 2 Pathology Department, Centro Hospitalar e Universit ario de Coimbra, Portugal 3 IBILI e Institute for Biomedical Imaging and Life Sciences, Faculty of Medicine, University of Coimbra j CIMAGO, Portugal 4 General Surgery Department, Unidade Local de Saúde de Castelo Branco, Portugal 5 CICS e Health Sciences Research Centre, Faculty of Health Sciences, University of Beira Interior, Portugal 6 rio de General Surgery Department, Centro Hospitalar e Universita Coimbra j IBILI j CIMAGO, Portugal