Functional Analysis of CCR4-NOT Complex in Pancreatic β Cell
Tu1759 A CD166 Negative Subpopulation of Pancreatic Cancer Cells Has Strong Invasive and Migratory...
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NF-κB, TNFα, IL-6 NOX2/4 (p ,0.001) were increased significantly. Conclusions: We suggest that the Sirt1/PGC1α/ Nrf1/Nrf2 pathways in liver may help to attenuate oxidative and inflammatory stress after RYGB procedure. Correcting the dysregulation of these mole- cules will benefit patients with obesity-induced metabolic syndromes. Tu1759 A CD166 Negative Subpopulation of Pancreatic Cancer Cells Has Strong Invasive and Migratory Activity Kenji Fujiwara, Kenoki Ohuchida, Koji Shindo, Daiki Eguchi, Shingo Kozono, Takao Ohtsuka, Shunichi Takahata, Shinichi Aishima, Kazuhiro Mizumoto, Masao Tanaka BACKGROUND: CD166 expression is correlated with prognosis in several cancers. However, its significance in pancreatic cancer is not clear. The aim of this study is to clarify the significance of CD166 expression in pancreatic cancer. METHODS: We performed flow cytometry to analyze expression of CD166 in pancreatic cancer cell lines. We also analyzed the functional differences between CD166+ and CD166- cells using invasion, migration and proliferation assays. We performed immunohistochemistry to investigate CD166 expression in surgically resected pancreatic cancer tissues. RESULTS: In flow cytometry, CD166 was expressed in pancreatic cancer cells in wide range (0-99.5%). In invasion assay, the invasive- ness of CD166- cells was greater than that of CD166+ cancer cells (p ,0.05). .In migration assay, the migratory activity of CD166- cells was greater than that of CD166+ cancer cells (p,0.05). In proliferation assay, there was no significant difference between CD166+ pancreatic cancer cells and CD166- cells. The analysis of real-time quantitative RT-PCR revealed that epithelial-mesenchymal transition activator Zeb1 mRNA was over-expressed in CD166- cells (p,0.001 compared with that in CD166 + cells). In immunohistochemistry, there was no significant difference in prognosis between CD166 high staining group (15 patients; 48.4%) and CD166 low staining group (16 patients; 51.6%). CONCLUSION: Our findings suggest that CD166- cells exhibit more aggressive behavior and activation of Zeb1 may play a role in this behavior, although further investigation is needed. Tu1760 Podoplanin Expressing Fibroblasts Enhance the Tumor Progression of Invasive Ductal Carcinoma of Pancreas, and Podoplanin Expression Was Affected by Cultured Condition Koji Shindo, Shinichi Aishima, Kenoki Ohuchida, Kazuhiro Mizumoto, Masao Tanaka, Yoshinao Oda Background: An interaction between cancer cells and surrounded cancer associated fibroblasts (CAFs) plays an important role in the progress of cancer. Pancreatic cancer is characterized by a growth of abundant fibrous or connective tissue, called "desmoplasia", and hypovascular environment inducing hypoxic and undernutritional condition. In pancreatic cancer, CD10+ myofibroblast-like activated Pancreatic Stellate Cells (PSCs) enhance the progression of pancreatic cancer by secreting high levels of MMP3 (Ikenaga, et al. Gastroenterology 2010). Podoplanin, usually used as a lymphatic vessels marker (D2-40), had been described as a predictor of prognosis in various types of cancer when it was expressed in involved stromal fibroblasts. Methods: We investigated Podoplanin expression in fibroblasts involved in pan- creatic cancerous tissue using immunohistochemistry (IHC). We established primary cultured fibroblasts as CAFs of fresh pancreatic adenocarcinoma tissue by out-growth method, and analyzed Podoplanin expression of CAFs using qRT-PCR and flow cytometry. We sorted CAFs by Magnetic Activated Cell Sorting (MACS) according to the expression of Podoplanin, and compared Podoplanin+ CAFs with Podoplanin- CAFs by migration assay and invasion assay in co-culture with pancreatic cancer cell lines. In addition, we performed qRT-PCR to elucidate differentiation between them. We also compared Podoplanin high-expressing CAFs with Podoplanin knocked down CAFs by siRNA to clarify the own function of Podoplanin. Next, we investigated the Podoplanin inducible condition by a time course of expression analysis using total starvation medium (EBSS), and DMEM added by recombinant growth factors or several percentages of FBS. Results: IHC showed that the frequency of Podoplanin expression (.30%) in fibroblasts was associated with lymphatic invasion, venous invasion, tumor size ( .3cm), histological grade, pT, and a shorter survival time (P ,0.001). Podoplanin expression in cultured CAFs showed heterogeneity (ranging from 0 to 95%) by flow cytometry. Podoplanin+ CAFs showed significantly high expression of CD10 and MMP3 compared with Podoplanin- CAFs, and co-culture experiments using sorted CAFs showed that Podoplanin+ CAFs enhanced the ability of cancer cells in migration and invasion compared with Podoplanin- CAFs (P ,0.05), while knock down of Podoplanin showed no effect on migration and invasion assay. Podoplanin expression in CAFs was up-regulated in the condition of starvation and lower concentration of growth factors or FBS. Conclusion: Despite Podoplanin in CAFs had no function of own to affect cancer cells, Podoplanin+ CAFs enhanced the progression of pancreatic cancer cells by co-expression of CD10 and MMP3. Podoplanin expression was up-regulated in the condition of starvation and lower concentration of growth factors or FBS. Tu2131 The Clinical Utility of the Local Inflammatory Response in Colorectal Cancer Colin H. Richards, Campbell S. Roxburgh, Arfon G. Powell, Alan K. Foulis, Paul G. Horgan, Donald C. McMillan Background The host immune response is important in the prevention of tumour progression in solid organ cancers but is not utilised in clinical practice. The aim was to evaluate the clinical utility of the local inflammatory response in patients with colorectal cancer. Methods Three hundred and sixty-five patients with primary operable colorectal cancer were included. The local inflammatory response was assessed using three different methods; (1) individual immune cells (CD3+, CD8+, CD45R0+, FOXP3+); (2) a composite immunohistochemistry- based score (Galon Immune Score); (3) a histopathological assessment (Klintrup-Makinen grade). Relationships with tumour and host characteristics were established and the prognos- tic value of each method compared. Results A strong infiltration of tumour infiltrating S-1135 SSAT Abstracts lymphoctyes (TIL's) was associated with improved cancer specific survival. When specific T-cell subtypes were considered, CD3+ was the strongest predictor of survival at both the invasive margin (CD3+ IM) and tumour stroma (CD3+ ST) while CD8+ was the strongest predictor in the cancer cell nests (CD8+ CCN). Infiltration of TIL's was associated with early tumour stage, an expanding growth pattern and lower levels of venous invasion but was not influenced by host characteristics or systemic inflammation. The Galon Immune Score and the Klintrup-Makinen grade were strongly related to individual T-cell infiltration and all three methods exhibited similar survival relationships in both node-positive and node- negative disease. Conclusion A coordinated adaptive immune response is an important factor in predicting outcome in patients with colorectal cancer. By comparing different methodologies we have provided a foundation on which to develop a standardised approach for assessing tumour inflammatory cell infiltrate. Figure 1. Kaplan-Meier survival curves demonstrating the cancer specific survival of patients with primary operable colorectal cancer according to the application of proposed immune scores. Clockwise from top left; CD+ IM, CD8+ CCN, K-M grade and the Galon Immune Score (strong to weak infiltration are shown top to bottom). Tu2132 Can Serum Lactate Predict the Outcome of Patients Who Underwent Emergent Exploratory Laparotomy for Acute Abdomen? Kaori Ito, Cheryl Anderson, Marc D. Basson Background: Serum lactate is a biomarker that predicts mortality in patients with non- cardiogenic circulatory shock like sepsis or severe trauma. Some reports suggest using the serum lactate to predict the prognosis in patients with acute abdomen, but this is not well understood. We hypothesized that the preoperative serum lactate level can help to predict the postoperative outcome in patients undergoing exploratory laparotomy for acute abdomen. Methods: Medical records of 293 consecutive patients who underwent emergent exploratory laparotomy for acute abdomen from 2007 through 2010 were reviewed. Patients' demograph- ics, preoperative laboratory tests including white blood cell counts (WBC), serum lactate, postoperative diagnosis, Systemic Inflammatory Response Syndrome (SIRS) Score, the Ameri- can Society of Anesthesiologists (ASA) physical status classification, postoperative in-hospital mortality were reviewed. These factors were compared between patient who died in hospital after the exploration and who survived, as well as between patients with bowel ischemia and without bowel ischemia. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) for the mortality and bowel ischemia were analyzed. PASW Statistics 18 was utilized for statistical analysis. Results: Two hundred and two patients who had recorded preoperative serum lactate(s) were included in the study. The preoperative serum lactate was checked only once in 130 patients and more than once in 72 patients. For patients with serial lactates, the trend over time was recorded. Postoperative diagnoses were small bowel obstruction (n=67, 33%), large bowel obstruction (n=44, 22%), bowel ischemia (n=38, 19%), perforated gastric or duodenal ulcers (n=31, 15%), colonic perforation (n=13, 6%), acute diverticulitis (n=19, 9%), others (n=21, 10%), and negative exploration (n=3, 1%). There were 34 (17%) postoperative in-hospital mortalities (The median time between surgery to death: 6.5 days [Range: 0 - 102]). All 3 patients who underwent negative laparotomy had a normal serum lactate(s) preoperatively. As shown on Table 1, the persistent abnormal or up-trending serum lactate was seen more frequently in patients in who died in hospital after exploration (53% vs 26%, p= 0.002); as well as, in patients with bowel ischemia (46% vs 28%, p=0.090) . The serum lactate had the similar specificity and NPV to SIRS score, ASA class and WBC; however, had the lower sensitivities than other factors. Conclusion: Normal or down-trending serum lactate strongly predicts postoperative survival in patients who undergo emergent exploratory laparotomy for acute abdomen, although persistently elevated serum lactate does not necessarily predict mortality. It may be useful prognostic information for patients and families if combined with other factors. Table 1 SSAT Abstracts
Transcript of Tu1759 A CD166 Negative Subpopulation of Pancreatic Cancer Cells Has Strong Invasive and Migratory...
NF-κB, TNFα, IL-6 NOX2/4 (p,0.001) were increased significantly. Conclusions: Wesuggest that the Sirt1/PGC1α/ Nrf1/Nrf2 pathways in liver may help to attenuate oxidativeand inflammatory stress after RYGB procedure. Correcting the dysregulation of these mole-cules will benefit patients with obesity-induced metabolic syndromes.
Tu1759
A CD166 Negative Subpopulation of Pancreatic Cancer Cells Has StrongInvasive and Migratory ActivityKenji Fujiwara, Kenoki Ohuchida, Koji Shindo, Daiki Eguchi, Shingo Kozono, TakaoOhtsuka, Shunichi Takahata, Shinichi Aishima, Kazuhiro Mizumoto, Masao Tanaka
BACKGROUND: CD166 expression is correlated with prognosis in several cancers. However,its significance in pancreatic cancer is not clear. The aim of this study is to clarify thesignificance of CD166 expression in pancreatic cancer. METHODS: We performed flowcytometry to analyze expression of CD166 in pancreatic cancer cell lines. We also analyzedthe functional differences between CD166+ and CD166- cells using invasion, migration andproliferation assays. We performed immunohistochemistry to investigate CD166 expressionin surgically resected pancreatic cancer tissues. RESULTS: In flow cytometry, CD166 wasexpressed in pancreatic cancer cells in wide range (0-99.5%). In invasion assay, the invasive-ness of CD166- cells was greater than that of CD166+ cancer cells (p ,0.05). .In migrationassay, the migratory activity of CD166- cells was greater than that of CD166+ cancercells (p,0.05). In proliferation assay, there was no significant difference between CD166+pancreatic cancer cells and CD166- cells. The analysis of real-time quantitative RT-PCRrevealed that epithelial-mesenchymal transition activator Zeb1 mRNA was over-expressedin CD166- cells (p,0.001 compared with that in CD166 + cells). In immunohistochemistry,there was no significant difference in prognosis between CD166 high staining group (15patients; 48.4%) and CD166 low staining group (16 patients; 51.6%). CONCLUSION: Ourfindings suggest that CD166- cells exhibit more aggressive behavior and activation of Zeb1may play a role in this behavior, although further investigation is needed.
Tu1760
Podoplanin Expressing Fibroblasts Enhance the Tumor Progression of InvasiveDuctal Carcinoma of Pancreas, and Podoplanin Expression Was Affected byCultured ConditionKoji Shindo, Shinichi Aishima, Kenoki Ohuchida, Kazuhiro Mizumoto, Masao Tanaka,Yoshinao Oda
Background: An interaction between cancer cells and surrounded cancer associated fibroblasts(CAFs) plays an important role in the progress of cancer. Pancreatic cancer is characterizedby a growth of abundant fibrous or connective tissue, called "desmoplasia", and hypovascularenvironment inducing hypoxic and undernutritional condition. In pancreatic cancer, CD10+myofibroblast-like activated Pancreatic Stellate Cells (PSCs) enhance the progression ofpancreatic cancer by secreting high levels of MMP3 (Ikenaga, et al. Gastroenterology 2010).Podoplanin, usually used as a lymphatic vessels marker (D2-40), had been described as apredictor of prognosis in various types of cancer when it was expressed in involved stromalfibroblasts. Methods: We investigated Podoplanin expression in fibroblasts involved in pan-creatic cancerous tissue using immunohistochemistry (IHC).We established primary culturedfibroblasts as CAFs of fresh pancreatic adenocarcinoma tissue by out-growth method, andanalyzed Podoplanin expression of CAFs using qRT-PCR and flow cytometry. We sortedCAFs by Magnetic Activated Cell Sorting (MACS) according to the expression of Podoplanin,and compared Podoplanin+ CAFs with Podoplanin- CAFs by migration assay and invasionassay in co-culture with pancreatic cancer cell lines. In addition, we performed qRT-PCRto elucidate differentiation between them. We also compared Podoplanin high-expressingCAFs with Podoplanin knocked down CAFs by siRNA to clarify the own function ofPodoplanin. Next, we investigated the Podoplanin inducible condition by a time course ofexpression analysis using total starvation medium (EBSS), and DMEM added by recombinantgrowth factors or several percentages of FBS. Results: IHC showed that the frequency ofPodoplanin expression (.30%) in fibroblasts was associated with lymphatic invasion, venousinvasion, tumor size (.3cm), histological grade, pT, and a shorter survival time (P,0.001).Podoplanin expression in cultured CAFs showed heterogeneity (ranging from 0 to 95%) byflow cytometry. Podoplanin+ CAFs showed significantly high expression of CD10 and MMP3compared with Podoplanin- CAFs, and co-culture experiments using sorted CAFs showedthat Podoplanin+ CAFs enhanced the ability of cancer cells in migration and invasioncompared with Podoplanin- CAFs (P,0.05), while knock down of Podoplanin showed noeffect on migration and invasion assay. Podoplanin expression in CAFs was up-regulatedin the condition of starvation and lower concentration of growth factors or FBS. Conclusion:Despite Podoplanin in CAFs had no function of own to affect cancer cells, Podoplanin+CAFs enhanced the progression of pancreatic cancer cells by co-expression of CD10 andMMP3. Podoplanin expression was up-regulated in the condition of starvation and lowerconcentration of growth factors or FBS.
Tu2131
The Clinical Utility of the Local Inflammatory Response in Colorectal CancerColin H. Richards, Campbell S. Roxburgh, Arfon G. Powell, Alan K. Foulis, Paul G.Horgan, Donald C. McMillan
Background The host immune response is important in the prevention of tumour progressionin solid organ cancers but is not utilised in clinical practice. The aim was to evaluate theclinical utility of the local inflammatory response in patients with colorectal cancer. MethodsThree hundred and sixty-five patients with primary operable colorectal cancer were included.The local inflammatory response was assessed using three different methods; (1) individualimmune cells (CD3+, CD8+, CD45R0+, FOXP3+); (2) a composite immunohistochemistry-based score (Galon Immune Score); (3) a histopathological assessment (Klintrup-Makinengrade). Relationships with tumour and host characteristics were established and the prognos-tic value of each method compared. Results A strong infiltration of tumour infiltrating
S-1135 SSAT Abstracts
lymphoctyes (TIL's) was associated with improved cancer specific survival. When specificT-cell subtypes were considered, CD3+ was the strongest predictor of survival at both theinvasive margin (CD3+ IM) and tumour stroma (CD3+ ST) while CD8+ was the strongestpredictor in the cancer cell nests (CD8+ CCN). Infiltration of TIL's was associated with earlytumour stage, an expanding growth pattern and lower levels of venous invasion but wasnot influenced by host characteristics or systemic inflammation. The Galon Immune Scoreand the Klintrup-Makinen grade were strongly related to individual T-cell infiltration andall three methods exhibited similar survival relationships in both node-positive and node-negative disease. Conclusion A coordinated adaptive immune response is an importantfactor in predicting outcome in patients with colorectal cancer. By comparing differentmethodologies we have provided a foundation on which to develop a standardised approachfor assessing tumour inflammatory cell infiltrate.
Figure 1. Kaplan-Meier survival curves demonstrating the cancer specific survival of patientswith primary operable colorectal cancer according to the application of proposed immunescores. Clockwise from top left; CD+ IM, CD8+ CCN, K-M grade and the Galon ImmuneScore (strong to weak infiltration are shown top to bottom).
Tu2132
Can Serum Lactate Predict the Outcome of Patients Who UnderwentEmergent Exploratory Laparotomy for Acute Abdomen?Kaori Ito, Cheryl Anderson, Marc D. Basson
Background: Serum lactate is a biomarker that predicts mortality in patients with non-cardiogenic circulatory shock like sepsis or severe trauma. Some reports suggest using theserum lactate to predict the prognosis in patients with acute abdomen, but this is not wellunderstood. We hypothesized that the preoperative serum lactate level can help to predictthe postoperative outcome in patients undergoing exploratory laparotomy for acute abdomen.Methods: Medical records of 293 consecutive patients who underwent emergent exploratorylaparotomy for acute abdomen from 2007 through 2010 were reviewed. Patients' demograph-ics, preoperative laboratory tests including white blood cell counts (WBC), serum lactate,postoperative diagnosis, Systemic Inflammatory Response Syndrome (SIRS) Score, the Ameri-can Society of Anesthesiologists (ASA) physical status classification, postoperative in-hospitalmortality were reviewed. These factors were compared between patient who died in hospitalafter the exploration and who survived, as well as between patients with bowel ischemiaand without bowel ischemia. Sensitivity, specificity, positive predictive value (PPV), andnegative predictive value (NPV) for the mortality and bowel ischemia were analyzed. PASWStatistics 18 was utilized for statistical analysis. Results: Two hundred and two patients whohad recorded preoperative serum lactate(s) were included in the study. The preoperativeserum lactate was checked only once in 130 patients and more than once in 72 patients.For patients with serial lactates, the trend over time was recorded. Postoperative diagnoseswere small bowel obstruction (n=67, 33%), large bowel obstruction (n=44, 22%), bowelischemia (n=38, 19%), perforated gastric or duodenal ulcers (n=31, 15%), colonic perforation(n=13, 6%), acute diverticulitis (n=19, 9%), others (n=21, 10%), and negative exploration(n=3, 1%). There were 34 (17%) postoperative in-hospital mortalities (The median timebetween surgery to death: 6.5 days [Range: 0 - 102]). All 3 patients who underwent negativelaparotomy had a normal serum lactate(s) preoperatively. As shown on Table 1, the persistentabnormal or up-trending serum lactate was seen more frequently in patients in who diedin hospital after exploration (53% vs 26%, p= 0.002); as well as, in patients with bowelischemia (46% vs 28%, p=0.090) . The serum lactate had the similar specificity and NPVto SIRS score, ASA class and WBC; however, had the lower sensitivities than other factors.Conclusion: Normal or down-trending serum lactate strongly predicts postoperative survivalin patients who undergo emergent exploratory laparotomy for acute abdomen, althoughpersistently elevated serum lactate does not necessarily predict mortality. It may be usefulprognostic information for patients and families if combined with other factors.Table 1
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