- Email: [email protected]
Su2066 Appendiceal Mucocele: Varied Presentations of an Obscure Diagnosis
fractionated maple syrup A based on molecular weight by ultrafiltration. Then, proteins in the high molecular fraction of maple syrup were isolated by ammonium sulfate precipitation method. Cell growth assay of CRC cells that were administered obtained fraction or componets of maple syrup was performed. Results: The number of apototic cells in CRC cells that were administered maple syrup A were not significantly different as compered to those were administered maple syrup B. A total of 715 proteins were identified, and 388 proteins expression level had changed by more than 1.5-fold as a result of administration of each maple syrup by semi-quantification based on spectral counting. These differentially expressed proteins were classified into cell cylce pathway by GO analysis. CRC cells that were administered fraction which has molecular weight more than 10,000 showed significantly lower growth rate as compared to control cells. Moreover, CRC cells that were administered protein components in maple syrup A showed significantly lower growth rate than those in maple syrup B. Conclusion: Protein component in maple syrup might inhibit cell proliferation to regulate cell cycle and be a novel anti-cancer medicine candidates.
Targeting HSP90 in Patient derived Barrett's Esophagus and Esophageal adenocarcinoma cells Prabhu Ramamoorthy, Sreekar Vennelaganti, Prashanth Vennalaganti, Abhiram Duvvuri, Neil Gupta, Prateek Sharma Background: Barrett's esophagus (BE) is a known complication of gastroesophageal reflux disease. Furthermore, BE is associated with an increased risk of developing esophageal adenocarcinoma (EAC). Hence, identifying ways to suppress BE is beneficial. In this regard, one of our long term goals is to determine the role of stem cells, the rare quiescent cells in a tissue on progression of BE. Heat shock protein 90 (Hsp90) is a chaperone that prevents protein aggregation. It has been demonstrated to specifically protect oncogenic variants of signaling molecules from degradation and hence may serve as a therapeutic target for the treatment of esophageal cancer or for preventing BE from progressing to EAC. Aims: To study the effect of three natural compounds, celastrol, triptolide (both from Chinese Thunder God of Vine, Tripterygium wilfordii) and Gedunin (from Indian Neem tree, Azadirachta indica) compounds that target HSP90 on patient derived Barrett's esophagus and esophageal cancer cells grown both in standard 2-dimensional (2D) culture system, and in 3-dimensional (3D) spheroid system. Methods: Cells were isolated from a BE and EAC patient and plated in a tissue culture dish in DMEM/F12 media containing 15% fetal bovine serum (FBS). Cell growth was determined by proliferation (hexosaminidase assay) and clonogenicity assays. To determine effects on stem cells, cells were grown in ultra-low attachment round-bottomed plates to form 3D spheroidal structures. Results: Esophageal cells were obtained from 2 male patients with histologically proven EAC (Eac-1, age 79 y, BE length 9 cm) and nondysplastic BE (Ebe1, age 75 y, BE length 12 cm). All three compounds affected the proliferation of the two cell lines in a dose dependent manner. IC50 value was calculated for al three compounds and was calculated to be 0.75-1.5 µM, 36-42 nM and 20-40 µM for Celastrol, Triptolide and Gedunin, respectively. We also performed clonogenicity assays, where the three cell lines were treated for 48 h following which they were allowed to grow and form colonies. While all three compounds were effective in inhibiting colony formation, celastrol and triptolide appeared to be the most effective. Interestingly, 3D culture studies showed very different results. While celastrol treatment showed significant reduction in spheroid numbers of Eca1 cells compared to Ebe1 cells, Triptolide had similar response in both cell lines. More importantly, Gedunin had no effect on the cells. Conclusion: Celastrol appears to be the best compound for suppressing EAC cells, while triptolide affects both BE and EAC. More importantly, the studies suggest that the compounds affect stem cells in both these cells. In addition, the data suggest that we can use the cells derived from patient tissue to determine efficacy of a compound.
Su2066 Appendiceal Mucocele: Varied Presentations of an Obscure Diagnosis Sujievvan Chandran, Gabor Kandel, Callum Dargavel, Niroshan Muwanwella, Tareq Alomani, Yuto Shimamura, Hamzah Akram, Paul P. Kortan, Jeffrey D. Mosko, Christopher W. Teshima, Gary R. May, Norman E. Marcon Introduction: Appendiceal mucoceles are a rare entity that may present with a wide spectrum of symptoms. Its recognition is important given the underlying neoplastic potential. The current literature is predominantly confined to case reports or small case series with a lack of long-term follow-up. Methods: A retrospective study was conducted at St. Michael's hospital, Toronto, Canada. The pathology database was searched for the terms "appendix" and "mucocele" from January 2010 to November 2015. Medical records were reviewed for patient demographics, presenting symptoms, endoscopic investigations, surgical intervention and complications that may have arisen on long term follow-up. Results: Twenty-five patients with an appendiceal mucocele were identified. Cohort demographics included a male to female ratio of 12:13 and median age of 57 years (range 24 - 78 years). Follow up was available for 22/25 (88%) patients over a median 9 months (range 1-58). Eleven (44%) patients were symptomatic with abdominal pain, which lead to further investigations that identified the mucocele. Of the remaining 14/25 (56%) patients who were asymptomatic, 6/14 (43%) had their mucocele identified at screening endoscopy, 5/14 (35.7%) were incidental findings on imaging and 3/14 (21.4%) at surgery for unrelated indications. Two of these had surgery for underlying inflammatory bowel disease. Fifteen (60%) patients had a colonoscopy prior to surgery, however only 8/15 (53.3%) had their mucocele identified. Twenty-one (84%) had CT imaging prior to surgery and 19/21 (90.5%) identified appendiceal pathology. Surgical intervention was predominantly via a laparotomy (13/25, 52%) during which an appendicectomy (13/25, 52%) was most commonly performed. The median size of the resected appendix was 7cm (range 2.3-12cm) with predominantly low-grade (23/ 25, 92%) histology, with one case (1/25, 4%) each respectively of focal high grade and adenocarcinoma. Surgical margins were clear in 21/25 (84%) specimens. No cases of pseudomyxoma peritonii were observed across our cohort on long-term follow-up and specifically on the subgroup with positive surgical margins. Conclusion: Appendiceal mucocele has a varied presentation. CT scan is more sensitive than colonoscopy. A high index of clinical suspicion is required for detection early enough for cure.
Su2064 Sildenafil Treatment Suppresses Intestinal Tumorigenesis in Mice Darren D. Browning, Bianca N. Islam, Sarah Sharman, Allison Bridges, Subbaramiah Sridhar The guanylin/GC-C/cGMP signaling axis controls homeostasis in the intestinal mucosa and has been implicated in the suppression of visceral pain, colitis, and colon cancer. We recently showed that treating mice with the PDE 5 inhibitor Vardenafil (Levitra™) increased cGMP in the colon mucosa, and resulted in reduced proliferation and suppression of colitis. In the present study we tested the hypothesis that sustained increases in intestinal cGMP using Sildenafil will suppress tumorigenesis in preclinical models of colon cancer. We found that Sildenafil provided ad libitum significantly reduced tumor multiplicity in both Apcmin and DSS/AOM models by 40-50% (respectively). The intestinal polyps from the Apcmin mice treated with Sildenafil were ostensibly indistinguishable from untreated tumors with respect to indices of proliferation, differentiation and apoptosis. In contrast, the colonic polyps from the DSS/AOM mice treated with Sildenafil were significantly less proliferative, exhibited increased differentiation, but showed no difference in apoptosis. As observed previously with Vardenafil, Sildenafil reduced the severity of DSS-induced inflammation in the AOM treated mice as assessed by change in body weight. While this observation suggests a suppressive role for cGMP in tumor promotion in this inflammatory carcinogenesis model, Sildenafil treatment also resulted in reduced inflammation in the tumors as measured by inflammatory cytokines. Taken together our results suggest that Sildenafil treatment may have therapeutic benefit for the chemoprevention of colorectal cancer.
Su2067 Distinguishing Colorectal Neoplasia Stained With Fluorescently Labelled Fatty Acid Using Confocal Laser Endomicroscopy Feihong Deng, Yuan Fang, Biao Nie Background & Aims: As the first hallmark of neoplasm, the Warburg effect contributes to tumour tracing. However, the secondary hallmark, increased de novo fatty acid synthesis (FASN), has received little attention. The current study evaluated the topical use of a fluorescently labelled fatty acid analogue (BODIPY-FA) to detect colorectal cancer (CRC) in mouse models and patients. Methods: Metabolic pathways were analysed by comparing CRC samples with matched normal samples. Then, colorectal neoplasms were topically stained by BODIPY-FA, and the fluorescent signals were quantified in neoplastic and non-neoplastic sites with confocal laser endomicroscopy (CLE) and fluorescence microscopy. Results: The expression levels of FA transporters FABP1, Caveolin-1 and CD36 in normal tissues were 1.8-, 2.5- and 2.3-fold higher, respectively, compared with adenocarcinoma due to metabolic reprogramming, as determined via Western blotting. The BODIPY-FA signal in carcinomas or colonic intraepithelial neoplasia (IN) from three CRC mouse models were significantly decreased versus normal epithelium, as determined with CLE and fluorescence microscopy. The signals at human non-neoplastic mucosa sites were approximately 1.6, 2 and 2.2 times greater than that of low-grade IN, high grade IN and cancerous sites, respectively, as determined with CLE. Furthermore, BODIPY-FA staining not only showed the arrangement of glands and cellular structures but also displayed the nuclei. In a double-blind trial,CLE images with BODIPY-FA received higher consistency (K=0.68, 0.43 respectively) and greater overall validity(74.65%, 55.88% respectively) than with intravenous fluorescein sodium when compared with histological diagnosis Conclusions: CRC uptakes less fatty acid than the non-neoplastic epithelium. The topical use of BODIPY-FA is a promising imaging approach for screening colorectal neoplasia with CLE.
Su2065 Protein Component in Maple Syrup Has a Potential to Develop Novel AntiCancer Drugs for Colorectal Cancer Tetsushi Yamamoto, Ryota Shiburo, Kuniko Mitamura, Atsushi Taga Background: Colorectal cancer (CRC) is a leading cause of death among cancer patients, and many patients are already in advanced stages when they are diagnosed. Thus, effective CRC therapy is necessary for saving CRC patients. Maple syrup is popular natural sweetener in the world. Maple syrup contains not only carbohydrate such as sucrose but also various components including organic acids, amino acids, vitamins, and phenolic compounds. Furthermore, recent studies have shown that phenolic compounds in maple syrup may possess various activities such as decreasing blood glucose level and an anti-cancer effect. We previously reported that administration of a dark color type of maple syrup (maple syrup A) to CRC cells showed suppression of cell proliferation and invasion via inhibition of Akt activation while there was no effect of a light color type of maple syrup (maple syrup B) administation to CRC cells. In this study, we parformed proteomic analysis to clarify the inhibitory effect of maple syrup A on cell proliferation, and we also performed identification of the active ingredients in maple syrup A. Method: First, we mesured the number of the apoptotic cells using flowcytometer. Next, we administered maple syrup A or maple syrup B to CRC cells, and extracted protein. Then, the extracted protein was digested by trypsin, and digestion products were analyzed by liquid chromatography/mass spectormetry (LC/ MS). Gene ontology (GO) analysis were performed using identified proteins for each molecular function, biological process, cellular component and KEGG pathway term. Next, we
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Targeting HSP90 in Patient derived Barrett's Esophagus and Esophageal adenocarcinoma cells Prabhu Ramamoorthy, Sreekar Vennelaganti, Prashanth Vennalaganti, Abhiram Duvvuri, Neil Gupta, Prateek Sharma Background: Barrett's esophagus (BE) is a known complication of gastroesophageal reflux disease. Furthermore, BE is associated with an increased risk of developing esophageal adenocarcinoma (EAC). Hence, identifying ways to suppress BE is beneficial. In this regard, one of our long term goals is to determine the role of stem cells, the rare quiescent cells in a tissue on progression of BE. Heat shock protein 90 (Hsp90) is a chaperone that prevents protein aggregation. It has been demonstrated to specifically protect oncogenic variants of signaling molecules from degradation and hence may serve as a therapeutic target for the treatment of esophageal cancer or for preventing BE from progressing to EAC. Aims: To study the effect of three natural compounds, celastrol, triptolide (both from Chinese Thunder God of Vine, Tripterygium wilfordii) and Gedunin (from Indian Neem tree, Azadirachta indica) compounds that target HSP90 on patient derived Barrett's esophagus and esophageal cancer cells grown both in standard 2-dimensional (2D) culture system, and in 3-dimensional (3D) spheroid system. Methods: Cells were isolated from a BE and EAC patient and plated in a tissue culture dish in DMEM/F12 media containing 15% fetal bovine serum (FBS). Cell growth was determined by proliferation (hexosaminidase assay) and clonogenicity assays. To determine effects on stem cells, cells were grown in ultra-low attachment round-bottomed plates to form 3D spheroidal structures. Results: Esophageal cells were obtained from 2 male patients with histologically proven EAC (Eac-1, age 79 y, BE length 9 cm) and nondysplastic BE (Ebe1, age 75 y, BE length 12 cm). All three compounds affected the proliferation of the two cell lines in a dose dependent manner. IC50 value was calculated for al three compounds and was calculated to be 0.75-1.5 µM, 36-42 nM and 20-40 µM for Celastrol, Triptolide and Gedunin, respectively. We also performed clonogenicity assays, where the three cell lines were treated for 48 h following which they were allowed to grow and form colonies. While all three compounds were effective in inhibiting colony formation, celastrol and triptolide appeared to be the most effective. Interestingly, 3D culture studies showed very different results. While celastrol treatment showed significant reduction in spheroid numbers of Eca1 cells compared to Ebe1 cells, Triptolide had similar response in both cell lines. More importantly, Gedunin had no effect on the cells. Conclusion: Celastrol appears to be the best compound for suppressing EAC cells, while triptolide affects both BE and EAC. More importantly, the studies suggest that the compounds affect stem cells in both these cells. In addition, the data suggest that we can use the cells derived from patient tissue to determine efficacy of a compound.
Su2066 Appendiceal Mucocele: Varied Presentations of an Obscure Diagnosis Sujievvan Chandran, Gabor Kandel, Callum Dargavel, Niroshan Muwanwella, Tareq Alomani, Yuto Shimamura, Hamzah Akram, Paul P. Kortan, Jeffrey D. Mosko, Christopher W. Teshima, Gary R. May, Norman E. Marcon Introduction: Appendiceal mucoceles are a rare entity that may present with a wide spectrum of symptoms. Its recognition is important given the underlying neoplastic potential. The current literature is predominantly confined to case reports or small case series with a lack of long-term follow-up. Methods: A retrospective study was conducted at St. Michael's hospital, Toronto, Canada. The pathology database was searched for the terms "appendix" and "mucocele" from January 2010 to November 2015. Medical records were reviewed for patient demographics, presenting symptoms, endoscopic investigations, surgical intervention and complications that may have arisen on long term follow-up. Results: Twenty-five patients with an appendiceal mucocele were identified. Cohort demographics included a male to female ratio of 12:13 and median age of 57 years (range 24 - 78 years). Follow up was available for 22/25 (88%) patients over a median 9 months (range 1-58). Eleven (44%) patients were symptomatic with abdominal pain, which lead to further investigations that identified the mucocele. Of the remaining 14/25 (56%) patients who were asymptomatic, 6/14 (43%) had their mucocele identified at screening endoscopy, 5/14 (35.7%) were incidental findings on imaging and 3/14 (21.4%) at surgery for unrelated indications. Two of these had surgery for underlying inflammatory bowel disease. Fifteen (60%) patients had a colonoscopy prior to surgery, however only 8/15 (53.3%) had their mucocele identified. Twenty-one (84%) had CT imaging prior to surgery and 19/21 (90.5%) identified appendiceal pathology. Surgical intervention was predominantly via a laparotomy (13/25, 52%) during which an appendicectomy (13/25, 52%) was most commonly performed. The median size of the resected appendix was 7cm (range 2.3-12cm) with predominantly low-grade (23/ 25, 92%) histology, with one case (1/25, 4%) each respectively of focal high grade and adenocarcinoma. Surgical margins were clear in 21/25 (84%) specimens. No cases of pseudomyxoma peritonii were observed across our cohort on long-term follow-up and specifically on the subgroup with positive surgical margins. Conclusion: Appendiceal mucocele has a varied presentation. CT scan is more sensitive than colonoscopy. A high index of clinical suspicion is required for detection early enough for cure.
Su2064 Sildenafil Treatment Suppresses Intestinal Tumorigenesis in Mice Darren D. Browning, Bianca N. Islam, Sarah Sharman, Allison Bridges, Subbaramiah Sridhar The guanylin/GC-C/cGMP signaling axis controls homeostasis in the intestinal mucosa and has been implicated in the suppression of visceral pain, colitis, and colon cancer. We recently showed that treating mice with the PDE 5 inhibitor Vardenafil (Levitra™) increased cGMP in the colon mucosa, and resulted in reduced proliferation and suppression of colitis. In the present study we tested the hypothesis that sustained increases in intestinal cGMP using Sildenafil will suppress tumorigenesis in preclinical models of colon cancer. We found that Sildenafil provided ad libitum significantly reduced tumor multiplicity in both Apcmin and DSS/AOM models by 40-50% (respectively). The intestinal polyps from the Apcmin mice treated with Sildenafil were ostensibly indistinguishable from untreated tumors with respect to indices of proliferation, differentiation and apoptosis. In contrast, the colonic polyps from the DSS/AOM mice treated with Sildenafil were significantly less proliferative, exhibited increased differentiation, but showed no difference in apoptosis. As observed previously with Vardenafil, Sildenafil reduced the severity of DSS-induced inflammation in the AOM treated mice as assessed by change in body weight. While this observation suggests a suppressive role for cGMP in tumor promotion in this inflammatory carcinogenesis model, Sildenafil treatment also resulted in reduced inflammation in the tumors as measured by inflammatory cytokines. Taken together our results suggest that Sildenafil treatment may have therapeutic benefit for the chemoprevention of colorectal cancer.
Su2067 Distinguishing Colorectal Neoplasia Stained With Fluorescently Labelled Fatty Acid Using Confocal Laser Endomicroscopy Feihong Deng, Yuan Fang, Biao Nie Background & Aims: As the first hallmark of neoplasm, the Warburg effect contributes to tumour tracing. However, the secondary hallmark, increased de novo fatty acid synthesis (FASN), has received little attention. The current study evaluated the topical use of a fluorescently labelled fatty acid analogue (BODIPY-FA) to detect colorectal cancer (CRC) in mouse models and patients. Methods: Metabolic pathways were analysed by comparing CRC samples with matched normal samples. Then, colorectal neoplasms were topically stained by BODIPY-FA, and the fluorescent signals were quantified in neoplastic and non-neoplastic sites with confocal laser endomicroscopy (CLE) and fluorescence microscopy. Results: The expression levels of FA transporters FABP1, Caveolin-1 and CD36 in normal tissues were 1.8-, 2.5- and 2.3-fold higher, respectively, compared with adenocarcinoma due to metabolic reprogramming, as determined via Western blotting. The BODIPY-FA signal in carcinomas or colonic intraepithelial neoplasia (IN) from three CRC mouse models were significantly decreased versus normal epithelium, as determined with CLE and fluorescence microscopy. The signals at human non-neoplastic mucosa sites were approximately 1.6, 2 and 2.2 times greater than that of low-grade IN, high grade IN and cancerous sites, respectively, as determined with CLE. Furthermore, BODIPY-FA staining not only showed the arrangement of glands and cellular structures but also displayed the nuclei. In a double-blind trial,CLE images with BODIPY-FA received higher consistency (K=0.68, 0.43 respectively) and greater overall validity(74.65%, 55.88% respectively) than with intravenous fluorescein sodium when compared with histological diagnosis Conclusions: CRC uptakes less fatty acid than the non-neoplastic epithelium. The topical use of BODIPY-FA is a promising imaging approach for screening colorectal neoplasia with CLE.
Su2065 Protein Component in Maple Syrup Has a Potential to Develop Novel AntiCancer Drugs for Colorectal Cancer Tetsushi Yamamoto, Ryota Shiburo, Kuniko Mitamura, Atsushi Taga Background: Colorectal cancer (CRC) is a leading cause of death among cancer patients, and many patients are already in advanced stages when they are diagnosed. Thus, effective CRC therapy is necessary for saving CRC patients. Maple syrup is popular natural sweetener in the world. Maple syrup contains not only carbohydrate such as sucrose but also various components including organic acids, amino acids, vitamins, and phenolic compounds. Furthermore, recent studies have shown that phenolic compounds in maple syrup may possess various activities such as decreasing blood glucose level and an anti-cancer effect. We previously reported that administration of a dark color type of maple syrup (maple syrup A) to CRC cells showed suppression of cell proliferation and invasion via inhibition of Akt activation while there was no effect of a light color type of maple syrup (maple syrup B) administation to CRC cells. In this study, we parformed proteomic analysis to clarify the inhibitory effect of maple syrup A on cell proliferation, and we also performed identification of the active ingredients in maple syrup A. Method: First, we mesured the number of the apoptotic cells using flowcytometer. Next, we administered maple syrup A or maple syrup B to CRC cells, and extracted protein. Then, the extracted protein was digested by trypsin, and digestion products were analyzed by liquid chromatography/mass spectormetry (LC/ MS). Gene ontology (GO) analysis were performed using identified proteins for each molecular function, biological process, cellular component and KEGG pathway term. Next, we
S-625
X : 10052$CH01 03-28-16 00:57:27 PDFd : 10052B : o
Page 625
AGA Abstracts
AGA Abstracts
Su2063