bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 736: Hypoxia-induced EGFR tyrosine kinase inhibitor resistance is associated with epithelial-mesenchymal transition in NSCLC
Authors: Lu Yuhong , Glazer Peter M.
Published: July 2016

The development of small molecule inhibitors specific for epidermal growth factor receptors (EGFR) with activating mutations has led to a dramatic shift in the treatment of non-small cell lung cancer (NSCLC) patients. Patients with NSCLC carrying such EGFR mutations often show prolonged responses when treated with first or second generation EGFR tyrosine kinase inhibitors (TKIs). However, most patients eventually develop resistance to EGFR TKIs. Hypoxia is a key micro-environmental stress in solid tumors that is associated with poor prognosis. Studies have linked hypoxia to therapy resistance, including radiation therapy and chemotherapy. In this study, we show that long-term, moderate hypoxia induces gefitinib resistance in the NSCLC cell line, HCC827, that harbors an activating EGFR mutation. The hypoxia-induced gefitinib resistance is associated with reduced BIM induction after gefitinib treatment when compared to normoxic HCC827 cells. In addition, in hypoxic HCC827 cells, gefitinib treatment induces N-cadherin expression, a mesenchymal marker, accompanied by downregulation of E-cadherin, an epithelial maker. These results suggest that epithelial-mesenchymal transition (EMT) may be involved in hypoxia induced gefitinib resistance. Wound healing migration assays further support that hypoxia induces HCC827 cell migration after gefitinib treatment. These results suggest that hypoxia may be a driving force for EGFR TKIs resistance potentially through EMT in NSCLC.

Citation Format: Yuhong Lu, Peter M. Glazer. Hypoxia-induced EGFR tyrosine kinase inhibitor resistance is associated with epithelial-mesenchymal transition in NSCLC. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 736.
Abstract 740: Silica promotes anchorage-independent growth of human bronchial cells via activation of NF-êB1 p50
Authors: Katabami Motoo , Kinoshita Ichiro , Shimizu Yasushi , Takeuchi Satoshi , Dosaka-Akita Hirotoshi
Published: July 2016

Although crystalline silica (hereafter silica) was judged as carcinogen by IARC, the mechanism of silica-induced carcinogenesis remains elusive. As limited studies have been reported to support the direct carcinogenic effect of silica on human lung epithelial cells, silica-induced inflammatory microenvironment may be postulated to facilitate lung neoplastic transformation. To test this hypothesis, we investigate the growth of immortalized human lung bronchial epithelial cell lines NL20, BEAS-2B and 16HBE14o-, following incubation with conditioned media collected 48h after exposed to silica (Silica CM) in each cell line. All cell lines promoted anchorage-independent growth in each Silica CM, compared with those in CM without silica exposure. All the bronchial cell lines in Silica CM increased DNA binding activity of NF-êB1 p50. After incubated with Silica CM, culture supernatants of NL20 and BEAS-2B kept higher concentration of EGF, and those of 16HBE14o- contained higher concentration of TNFá and IGF1, respectively. Recombinant EGF, TNFá and IGF1 are all sufficient to promote anchorage-independent growth of each bronchial cell line, and to increase DNA binding activity of NF-êB1 p50. Neutralizing human EGF and TNFá antibodies dampened the growth of each bronchial cell line. NF-êB inhibitor Bay11-7082 suppressed the growth of all bronchial cell lines incubated with Silica CM. Taken together, silica promoted anchorage-independent growth of human lung epithelial cells via silica-induced secretion of soluble factors, followed by activation of NF-êB1 p50 in an autocrine manner. The present study supports the hypothesis that silica-induced inflammatory microenvironment may be actively involved in the potential carcinogenic effect of silica.

Citation Format: Motoo Katabami, Ichiro Kinoshita, Yasushi Shimizu, Satoshi Takeuchi, Hirotoshi Dosaka-Akita. Silica promotes anchorage-independent growth of human bronchial cells via activation of NF-êB1 p50. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 740.
Abstract 758: Modulating function of tumor associated macrophages by CSF1R inhibitor suppressed tumor growth in hepatocellular carcinoma
Authors: Sun Hui-Chuan , Ao Jian-Yang , Cai Hao , Chai Zong-Tao , Zhu Xiao-Dong
Published: July 2016

Aims: To investigate the effect of colony stimulating factor 1 receptor (CSF1R) inhibitor on macrophages and tumor pathogenesis.

Methods: CSF1 and CSF2 stimulated macrophages were obtained from mouse bone marrow derived monocytes cultured with CSF1 and CSF2. Mouse hepatoma cell line Hepa 1-6, and human hepatoma cell lines, HCCLM3 and HepG2 were used to established a allograft or xenograft tumor models to examine the effect of CSF1R inhibitor on tumor growth and distribution of macrophages. The characteristics of macrophages were determined by flow cytometry, immunohistochemistry, cell motility and Western blotting assays.

Results: CSF1R inhibitor suppressed tumor growth in the tumor models derived from three hepatoma cell lines, and prolonged hosts’ survival, but CSF1R inhibitor did not decreased the number of macrophages in Hepa1-6 allograft tumors. in vitro study showed CSF1R inhibitor suppressed proliferation and motility of CSF1 stimulated macrophages but not CSF2 stimulated macrophages. Furthermore, we found CSF1R inhibitor treatment decreased the ability of THP-1 macrophages to promote migration and invasion of tumor cells.

Conclusions: CSF1R inhibitor suppressed tumor growth in hepatoma animal models by modulating function of tumor associating macrophages.

Citation Format: Hui-Chuan Sun, Jian-Yang Ao, Hao Cai, Zong-Tao Chai, Xiao-Dong Zhu. Modulating function of tumor associated macrophages by CSF1R inhibitor suppressed tumor growth in hepatocellular carcinoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 758.
Abstract 766: Control of lung cancer with aspirin-triggered stimulation of resolution
Authors: Gilligan Molly M. , Sulciner Megan L. , Gus-Brautbar Yael , Ramon Sesquile , Colas Romain A. , Huang Sui , Serhan Charles N. , Panigrahy Dipak
Published: July 2016

Objective: To establish aspirin-triggered resolvins as a novel treatment to enhance current cancer therapies through the resolution of inflammation and clearance of cancer therapy-induced tumor cell debris. Background: Inflammation in the tumor microenvironment is now recognized as a strong promoter of tumor growth. Substantial epidemiological evidence suggests that aspirin, which suppresses inflammation, also reduces the risk of cancer. However, the mechanism by which aspirin reduces cancer risk and improves survival remains unclear, and toxicity has limited its clinical use. Low-dose aspirin stimulates the endogenous production of aspirin-triggered (AT) resolvins, novel omega-3 fatty acid-derived pro-resolving lipid autacoid mediators. We recently discovered that resolvins have potent anti-tumor activity, making resolvins potential candidates to mediate the anti-cancer activity of aspirin. Thus, we hypothesize that aspirin's anti-cancer activity is mediated in part by AT-resolvins via novel anti-inflammatory and pro-resolution mechanisms. Methods: Flow cytometry confirmed apoptotic and necrotic lung tumor cell debris following chemotherapy (etoposide) or targeted therapy (erlotinib) treatment in vitro. Macrophage phagocytosis was quantified via Relative Fluorescent Units (RFU). Cytokine levels were quantified via ELISA. Results: AT-resolvins counter therapy-induced debris-stimulated tumor growth and spontaneous lung metastases via enhancing endogenous clearance of tumor debris via macrophages and counter-regulating macrophage cytokine production. Pharmacological and genetic antagonism of the AT-resolvin D1 receptor (ALX/FPR2) neutralized aspirin's anti-cancer activity. Conclusions: AT-resolvins may provide a novel pathway and molecular mechanism to explain how aspirin reduces cancer risk so broadly. These anti-inflammatory and pro-resolving lipid autacoid mediators may complement current therapies for lung cancer with minimal toxicity.

Citation Format: Molly M. Gilligan, Megan L. Sulciner, Yael Gus-Brautbar, Sesquile Ramon, Romain A. Colas, Sui Huang, Charles N. Serhan, Dipak Panigrahy. Control of lung cancer with aspirin-triggered stimulation of resolution. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 766.
Abstract 769: Garcinol inhibits cancer stem cell-like phenotype via suppression of the Wnt/â-catenin /STAT3 axis signalling pathway in human non-small cell lung carcinomas
Authors: Huang. Wen-Chien
Published: July 2016

Innate or acquired drug resistance and consequently, tumor relapse in lung cancer patients have been linked to the activities of cancer stem cells (CSCs). Therefore, targeting CSCs is suggested as an effective approach for non-small cell lung cancer (NSCLC) therapy. In this study, we demonstrated that Garcinol, a polyisoprenylated benzophenone isolated from the fruiting bodies of Garcinia indica, possessing anti-inflammatory, antioxidant, acetyltransferase inhibitory and anticancer activities, modulates the activities of lung cancer stem cells (LCSCs) and their associated aggressiveness. Here, we demonstrated the inhibitory effect of Garcinol on LCSC phenotype of human NSCLC cells by using the analytical drug cytotoxicity or cell viability, flow-cytometric and functional assay approach. Garcinol significantly diminished the ability of NSCLC cell lines, H441 and A549 to form spheres. In parallel assays, Garcinol inhibited differentiated lung cancer cell and LCSCs viability in a dose-dependent manner. Consistent with these observations, Flow cytometric data showed that Garcinol reduced putative LCSC pool, evidenced by the dose-dependent decreased proportion of side-population (SP) cells and associated ALDH activity in Garcinol-treated H441 cells, compared to the control group. Additionally, functional assays showed that Garcinol markedly diminished the ability of H441 and A549 cells to form colonies. Mechanistically, Garcinol impaired phosphorylation of LRP6, a co-receptor of Wnt and STAT3. In same assay, Garcinol downregulated â-catenin, Dvl2, Axin2, and Cyclin D1 expressions in NSCLC-generated spheres, suggesting its ability to regulate the Wnt/â-catenin signalling pathway. Put together, we demonstrated here that Garcinol modulates the LCSC phenotype via regulation of the Wnt/â-catenin signalling and inactivation of STAT3, thus, presenting Garcinol as a putative novel anti-LCSC therapeutic agent.

Keywords - Garcinol, non-small cell lung cancer, cancer stem cells, Wnt/â-catenin signalling pathway, anti-cancer therapy, STAT3

Citation Format: Wen-Chien Huang. Garcinol inhibits cancer stem cell-like phenotype via suppression of the Wnt/â-catenin /STAT3 axis signalling pathway in human non-small cell lung carcinomas. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 769.
Abstract 774: Proteogenomic characterization of high-grade serous ovarian cancer
Authors: McDermott Jason E. , Payne Samuel , Ray Debjit , Petyuk Vladislav , Moore Ronald , Gritsenko Marina , Smith Richard , Rodland Karin
Published: July 2016

High grade serous ovarian carcinoma (HGSOC) is a highly lethal gynecological malignancy, in large part because most patients develop resistance to the standard of care chemotherapy with platinum and taxanes. We performed deep proteomic and phosphoproteomic characterization on ovarian high-grade serous carcinomas (HGSC) previously characterized genomically by the The Cancer Genome Atlas (TCGA) as part of the Clinical Proteomic Tumor Analysis Consortium (CPTAC; http://proteomics.cancer.gov/programs/cptacnetwork). We constructed an integrated proteogenomic landscape of HGSC and identified functional modules statistically associated with outcomes including platinum resistance and overall survival.

Integration of genomic data such as copy number alterations (CNA) with global protein abundance data revealed several chromosomal regions that have significant trans effects on protein expression. These genetically affected proteins were used to construct statistical models capable of predicting survival outcomes. Trans-affected proteins were enriched in proliferation, cell motility and invasion, and immune regulation, hallmarks of cancer.

We used statistical methods to identify functional pathways able to discriminate between short surviving and long surviving patients. We found that protein abundance and phosphorylation indicated pathway activity that was able to clearly discriminate between these two groups, but that transcriptional expression and CNAs were not. We integrated proteomic abundance and phosphorylation state to elucidate specific signaling interactions and pathways differentially active in tumors from patients with short overall survival as well as to predict novel kinase substrate relationships important in HGSC. Finally, we identified druggable pathways that are differentially active across proteomic subtypes.

A key point arising from our multimodal analysis is that combining different types of molecular data from these tumors greatly increases the ability to identify biologically relevant differences between groups. Using a Lasso-based regression approach to integrate data we show that the most robustly predictive survival models can be obtained using integrated CNA, transcriptome, proteome, and phosphoproteome data than with any of the individual sources of data.

Citation Format: Jason E. McDermott, Samuel Payne, Debjit Ray, Vladislav Petyuk, Ronald Moore, Marina Gritsenko, Richard Smith, Karin Rodland. Proteogenomic characterization of high-grade serous ovarian cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 774.
Abstract 777: The CoGAPS matrix factorization algorithm infers feedback mechanisms from therapeutic inhibition of EGFR that increases expression of growth factor receptors
Authors: Fertig Elana J. , Ozawa Hiroyuki , Thakar Manjusha , Howard Jason , Krigsfeld Gabriel , Favorov Alexander V. , Gaykalova Daria A. , Ochs Michael F. , Chung Christine H.
Published: July 2016

Next generation sequencing technologies enable precise personalized medicine. Thus, patients with oncogene driven tumors are currently treated with targeted therapeutics such as EGFR inhibitors. However, drug interactions with other activated signaling pathways in treated tumors often alter predicted therapeutic response. Therefore, bioinformatics algorithms are needed to infer unanticipated molecular interactions from anticipated molecular response to targeted therapeutics in diverse genetic backgrounds. To model heterogeneous genetic backgrounds in HNSCC, we use HaCaT cells with forced overexpression of EGFR, HRAS, and PIK3CA. Previously, the CoGAPS matrix factorization algorithm was shown to infer the specific signaling pathways that were activated in these HaCaT knock-in constructs from gene expression data. In this study, we evaluated whether CoGAPS could also delineate unanticipated signaling changes from anticipated cellular signaling response caused by targeted therapeutic in diverse genetic backgrounds. To delineate these signaling responses, we measured gene expression after treating the modified HaCaT cells with three EGFR targeted agents (gefitinib, cetuximab and afatinib) for 24 hours. The CoGAPS matrix factorization algorithm distinguished a gene expression signature associated with the anticipated silencing of the EGFR network and a signature associated with unanticipated transcriptional feedback in HaCaT constructs that were sensitive to EGFR inhibitors. Notably, the feedback signature showed that EGFR gene expression itself increased in cells that were responsive to EGFR inhibitors. The CoGAPS algorithm further associated such feedback with increased expression of several growth factor receptors by the AP-2 family of transcription factors. Once transcribed, these growth factor receptors may ultimately compensate for EGFR inhibition in these sensitive cells. Our data suggest, that CoGAPS gene expression signatures delineate on target and feedback effects of drugs related to therapeutic sensitivity in diverse genetic backgrounds.

Citation Format: Elana J. Fertig, Hiroyuki Ozawa, Manjusha Thakar, Jason Howard, Gabriel Krigsfeld, Alexander V. Favorov, Daria A. Gaykalova, Michael F. Ochs, Christine H. Chung. The CoGAPS matrix factorization algorithm infers feedback mechanisms from therapeutic inhibition of EGFR that increases expression of growth factor receptors. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 777.
Abstract 79: Andrographolide alters genes associated with DNA repair in prostate cancer
Authors: Forestier-Román Ingrid , Sánchez-Vázquez María , Rohena-Rivera Krizia , Cadilla Carmen , Martínez-Ferrer Magaly
Published: July 2016

Prostate cancer is the most frequently diagnosed non-cutaneous cancer and the second cause of cancer-related deaths in American men. Andrographolide, a labdane diterpenoid that is a component of the medicinal plant Andrographis paniculata, has been reported to have a wide range of biological activities including anticarcinogenic properties. In previous studies, we found that Andrographolide inhibits cell growth, tumor growth and angiogenesis in prostate cancer. Therefore, the objective of this study is to determine the mechanism of action of Andrographolide in prostate cancer. In order to determine the mechanism of action, we did gene expression profile to compare genes differentially expressed in tumors treated with Andrographolide (10 mg/kg and 25 mg/kg) and their vehicle. Tumors were developed using a xenograft model in which the prostates of SCID mice were injected with 22RV1, and mice were treated bi-weekly with Andrographolide (10 mg/kg and 25 mg/kg). Tumor tissues were collected and snap frozen. Gene expression was analyzed using the Affymetrix GeneChip® Human Gene 2.0 array. Microarray studies showed a total of 674 genes differentially expressed in tumors treated with Andrographolide (10 mg/kg) when compared to their vehicle. In addition, 218 genes were differentially expressed in tumors treated with Andrographolide (25 mg/kg) when compared to their vehicle. Genes involved in cellular processes such as cell cycle, DNA damage response, MAPK signaling, TCA cycle, glycolysis and metabolism of amino acids were differentially expressed. Ingenuity Pathway Analysis (IPA) revealed networks associated to DNA replication, recombination and repair, and post-translational modification in tumors treated with Andrographolide (10 mg/kg). Moreover, IPA revealed networks associated to cell death and survival, cellular function and maintenance, and carbohydrate metabolism in tumors treated with Andrographolide (25 mg/kg). The microarray results were confirmed by quantitative real-time PCR. Our results demonstrated that Andrographolide altered the expression of genes associated with DNA repair which could be a possible mechanism of action of its anticarcinogenic effect.

Citation Format: Ingrid Forestier-Román, María Sánchez-Vázquez, Krizia Rohena-Rivera, Carmen Cadilla, Magaly Martínez-Ferrer. Andrographolide alters genes associated with DNA repair in prostate cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 79.
Abstract 800: ALDH2 rs671 polymorphism and alcohol consumption in the risk of gastric cancer: A Mendelian randomization study
Authors: Kim Jeongseon , Yang Sarah , Lee Jeonghee , Choi Il Ju , Kim Young Woo , Ryu Keun Won , Sung Joohon
Published: July 2016

Background: The effect of increased alcohol consumption on the risk for gastric cancer (GC) has not been fully elucidated, due to the presence of confounders and gender differences. In this study, aldehyde dehydrogenase 2 (ALDH2) rs671, the genetic polymorphism related to the elimination of acetaldehyde, was used as an instrumental variable (IV) for Mendelian randomization analysis to dissect the role of alcohol in GC risk.

Methods: The study included Korean 450 GC cases and 1,050 controls recruited at the National Cancer Center. Data of 795 GC patients and 4,893 controls from the Korean Centers for Disease Control were used for further validation. The odds ratios (OR) for the ALDH2 genotype and its interaction with alcohol consumption were estimated using multivariate logistic regression. The two-stage control function method was performed using rs671 as the IV for alcohol consumption.

Results: All AG carriers had an OR of 1.26 (95% confidence interval, 1.11-1.42) compared with GG carriers. In a quantitative analysis of alcohol intake, the conventional OR was 1.21(1.09-1.36) and the IV-OR was 0.90(0.63-1.28). Females had low alcohol consumption and did not show a significant risk for GC while males had a high risk for GC.

Conclusions: Alcohol consumption is not associated with a risk for GC by itself, and ALDH2 polymorphism should be considered simultaneously when assessing the risk. Females did not display an evident risk for GC regarding alcohol, partly due to their lower level of alcohol consumption. However, further dissection of gender difference is necessary.

Citation Format: Jeongseon Kim, Sarah Yang, Jeonghee Lee, Il Ju Choi, Young Woo Kim, Keun Won Ryu, Joohon Sung. ALDH2 rs671 polymorphism and alcohol consumption in the risk of gastric cancer: A Mendelian randomization study. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 800.
Abstract 803: Obesity and associated lifestyles modify the effect of glucose metabolism-related genetic variants on impaired glucose homeostasis among postmenopausal women
Authors: Jung Su Yon , Sobel Eric M. , Papp Jeanette C. , Crandall Carolyn J. , Fu Alan N. , Zhang Zuo-Feng
Published: July 2016

Background: Impaired glucose metabolism-related genetic variants likely interact with obesity-modifiable factors in response to glucose intolerance, yet their interconnected pathways have not been fully characterized. We assessed in postmenopausal women whether obesity, physical activity, and high dietary fat intake interact with the single-nucleotide polymorphisms (SNPs)-glucose variations.

Methods: We used data from 1,027 postmenopausal participants of the Genomics and Randomized Trials Network study and 15 SNPs associated with glucose homeostasis. We used regression analysis plus stratification and graphic approaches.

Results: Fasting levels of glucose, insulin, and homeostatic model assessment-insulin resistance (HOMA-IR) were higher in obese, inactive, and high fat-diet women than in their respective counterparts. Carriers within subgroups differently demonstrated the direction and/or magnitude of the variants’ effect on glucose-relevant traits. Variants in GCKR, GCK, DGKB/TMEM195 (P for interactions = 0.02, 0.02, and 0.01), especially, showed interactions with obesity: obese, inactive, and high fat-diet women had greater increases in fasting glucose, insulin, and HOMA-IR levels. Obese carriers at TCF7L2 variant had greater increases in fasting glucose levels than non-obese carriers (P for interaction = 0.04), whereas active women had greater decreases in insulin and HOMA-IR levels than inactive women (P for interaction = 0.02 in both levels).

Conclusions: Our data support the important role of obesity in modifying glucose homeostasis in response to glucose metabolism-relevant variants. These findings may inform research on the role of glucose homeostasis in the etiology of disease, particularly obesity-related cancer, and the development of intervention strategies to reduce risk in postmenopausal women.

Citation Format: Su Yon Jung, Eric M. Sobel, Jeanette C. Papp, Carolyn J. Crandall, Alan N. Fu, Zuo-Feng Zhang. Obesity and associated lifestyles modify the effect of glucose metabolism-related genetic variants on impaired glucose homeostasis among postmenopausal women. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 803.
Abstract 804: Gene-environment interactions and the risk of Barrett's esophagus in three US cohorts
Authors: Crous-Bou Marta , Feskanich Diane , Stampfer Meir J. , Fuchs Charles S. , De Vivo Immaculata , Jacobson Brian C.
Published: July 2016

Background: Several single nucleotide polymorphisms (SNPs) have been associated with Barrett's esophagus (BE) risk. Additionally, environmental factors including smoking, alcohol consumption and heartburn have been shown to increase BE risk. However, no studies have evaluated whether interactions between these genetic and environmental factors influence BE risk. Understanding how genes and environmental risk factors interact may provide key insight into the pathophysiology of BE.

Objectives: To examine the main effects and the potential effect-modification between known genetic loci (SNPS) and established environmental risk factors for BE.

Patients and Methods: We performed a nested case-control study using data on 401 incident BE cases and 436 age-matched controls from the Nurses’ Health Study, Nurses’ Health Study II, and Health Professionals Follow-up Study cohorts, who gave blood and completed biennial questionnaires. We genotyped SNPs identified in previous BE GWAS as well as SNPs in candidate genes related to BE susceptibility (i.e., related to excess body fat, fat distribution, factors associated with insulin resistance, and inflammatory mediators). A genetic risk score (GRS) was constructed to evaluate the combined effect of the selected SNPs on BE risk. Interactions between SNPs and BE risk factors were also assessed.

Results and Conclusions: We found a borderline association between our GRS and BE risk: a one-allele increase in the unweighted GRS increased the risk of Barrett's esophagus by a factor of 1.20 (95%CI = 1.00-1.44; p = 0.057). We observed a significant interaction between smoking and BE genotypes (p-interaction = 0.016). However, while alcohol consumption and heartburn duration were strongly associated with BE, we did not observe any significant gene-environment interactions.

Citation Format: Marta Crous-Bou, Diane Feskanich, Meir J. Stampfer, Charles S. Fuchs, Immaculata De Vivo, Brian C. Jacobson. Gene-environment interactions and the risk of Barrett's esophagus in three US cohorts. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 804.
Abstract 809: Dietary iron intake, genetic variants in microRNA related iron regulatory pathway genes, and the risk of non-small cell lung cancer
Authors: Zhang Liren , Ye Yuanquing , Tu Huakang , Hildebrandt Michelle A.t. , Heymach John V. , Roth Jack A. , Gu Jian , Wu Xifeng
Published: July 2016

As an essential nutrient that facilitates DNA synthesis, cell proliferation and metabolism, iron is involved in carcinogenesis. While epidemiological studies suggest a J-shaped model for the relationship of serum iron level and cancer risk with both too low and too high levels associated with increased cancer risks, the effect of genetic variants related to intracellular iron regulatory pathway and dietary iron intake on non-small cell lung cancer (NSCLC) risk is little known. In this study, we evaluated a panel of miRNA related genetic variants in iron regulatory pathway genes on the risk of NSCLC. In discovery phrase, 157 miRNA related single nucleotide polymorphisms (SNPs) from 81 genes of iron regulatory pathway were genotyped in 1656 NSCLC cases and 1486 healthy controls. Eight SNPs in 6 genes were significantly associated with risk of NSCLC. A SNP in 3’ UTR of Iron-Responsive Element Binding Protein 2 (IREB2) gene at 15q25 region was subsequently replicated in dbGaP lung cancer GWAS dataset. Combining all subjects (7,352 cases, 7,296 controls), the overall P-value was 4.9 × 10−9 (Odds ratio (OR) = 0.86). eQTL analysis showed that the SNP on IREB2 alters IREB2 gene expression (P = 3.0 × 10−11). The SNP is predicted to alter a miR-29 binding site on IREB2 gene and indeed the expression of miR-29 is inversely correlated with IREB2 expression in tumor tissues. We then determined the expression of serum miR-29a in 150 early-stage NSCLC patients and 172 healthy controls and found that miR-29a was significantly associated with higher risk of cancer (OR = 1.78, P = 0.03). By analyzing dietary intake information of the discovery population, iron intake was significantly associated with risk of NSCLC in logistic regression analysis and there was also evidence of a synergistic interaction with between the SNP and iron intake in modulating NSCLC risk (P for interaction = 0.01). Taken together, SNPs at miRNA binding sites of iron regulatory pathway genes and dietary iron intake may modify risk of NSCLC individually and jointly.

Citation Format: Liren Zhang, Yuanquing Ye, Huakang Tu, Michelle A.t. Hildebrandt, John V. Heymach, Jack A. Roth, Jian Gu, Xifeng Wu. Dietary iron intake, genetic variants in microRNA related iron regulatory pathway genes, and the risk of non-small cell lung cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 809.
Abstract 810: The CARRIERS consortium: Establishing refined breast cancer risk estimates in known predisposition genes
Authors: Lilyquist Jenna , Kraft Peter , Hart Steven N. , Hallberg Emily J. , Hu Chunling , Moore Raymond , Gnanaolivu Rohan , Domchek Susan M. , Weitzel Jeffrey N. , Nathanson Katherine L. , Goldgar David E. , Couch Fergus J.
Published: July 2016

Breast cancer is the most common type of cancer, and has a strong heritable component, with approximately 15 percent of new cases having family history of breast cancer. Genetic panel testing for patients with a family history of breast cancer has become a popular clinical resource. However, the risk of breast cancer associated with inactivating mutations in the predisposition genes is largely undefined, creating critical limitations in the interpretations of the results from panel testing and subsequent medical management. The Cancer Risk Estimates Related to Susceptibility Genes (CARRIERS) consortium was established to determine the risk of cancers associated with truncating mutations in the known predisposition genes and to extend these studies to the clinical classification of variants of uncertain significance (VUS) in the relevant genes.

CARRIERS involves screening of approximately 30,000 population-based breast cancer cases and 30,000 study matched controls for mutations in 28 established or proposed breast cancer predisposition genes. Population-based risks for breast cancer associated with mutations in each gene will be estimated. In parallel, 10,000 breast cancer cases from moderate and high-risk breast cancer families will be similarly screened for mutations and the penetrance of the mutations in high-risk families will be estimated. Furthermore, VUS identified by mutation screening will be characterized by functional and family-based studies and models for classification of the clinical relevance of VUS in each gene will be developed. These data are expected to lead to improvements in cancer risk assessment and improved management of patients found to carry mutations or VUS in breast cancer predisposition.

Citation Format: Jenna Lilyquist, Peter Kraft, Steven N. Hart, Emily J. Hallberg, Chunling Hu, Raymond Moore, Rohan Gnanaolivu, Susan M. Domchek, Jeffrey N. Weitzel, Katherine L. Nathanson, David E. Goldgar, Fergus J. Couch. The CARRIERS consortium: Establishing refined breast cancer risk estimates in known predisposition genes. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 810.
Abstract 822: Role of c-Myc in prostate cancer stem-like cell inhibition by sulforaphane
Authors: Kim Su-Hyeong , Vyas Avani R. , Singh Shivendra V.
Published: July 2016

Chemopreventive efficacy of D,L-sulforaphane (SFN) against prostate cancer has been shown in a transgenic mouse model but the underlying mechanism is not fully understood. The present study demonstrates that c-Myc, which is an oncogenic transcription factor frequently upregulated in human prostate cancer, is a molecular target of SFN. Exposure of human prostate cancer cells (LNCaP, C4-2, and PC-3) and a cell line derived from prostate adenocarcinoma of a transgenic mouse (Myc-CaP) to SFN resulted in a marked decrease in c-Myc protein level. Naturally-occurring thio, sulfinyl, and sulfonyl analogues of SFN were also effective in causing suppression of c-Myc protein level. The c-Myc is a known regulator of cellular metabolism but basal glycolysis was not altered by SFN treatment in empty vector transfected or in c-Myc overexpressing PC-3 cells. The effect of SFN treatment on prostate cancer stem-like cells (pCSC) was also determined as c-Myc is implicated in their maintenance. Analysis of aldehyde dehydrogenase 1 activity, CD49f+ fraction, and sphere forming efficiency revealed dose-dependent inhibition of pCSC in SFN-treated cells. The pCSC inhibition by SFN was partially but significantly attenuated by overexpression of c-Myc. Expression profiling revealed upregulation of a set of stemness genes by c-Myc overexpression but their downregulation by SFN treatment. Finally, oral administration of SFN to Transgenic Adenocarcinoma of Mouse Prostate (TRAMP) mice resulted in a marked decrease in c-Myc protein expression in the adenocarcinoma. In conclusion, this study indicates that c-Myc downregulation contributes to pCSC inhibition by SFN. This investigation was supported by grant CA115498 awarded by the National Cancer Institute.

Citation Format: Su-Hyeong Kim, Avani R. Vyas, Shivendra V. Singh. Role of c-Myc in prostate cancer stem-like cell inhibition by sulforaphane. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 822.
Abstract 824: Effects of metfornin on mammary carcinogenesis and metabolic profiles in methylnitrosourea-treated rats on standard or Western diets
Authors: Miller Mark S. , Thompson Matthew D. , Lubet Ronald A. , Steele Vernon E. , Seifried Harold E. , Grubbs Clinton J.
Published: July 2016

The methylnitrosourea-induced model of ER+ mammary cancer in female Sprague-Dawley rats has been routinely used to screen for chemopreventive agents. We recently reported that metformin was ineffective in reducing cancer formation in rats on a standard (4% fat) diet (Thompson, et al., Cancer Prev Res. 8, 231, 2015). In the present studies, rats were placed on a standard (Teklad) diet or on a Western diet (20% fat, low calcium) at 43 days of age (DOA), and administered MNU at 50 DOA. Rats were administered metformin or vehicle daily by gavage beginning at 57 days of age. Serum from the rats of the various groups was obtained both at 21 days following initial administration of metformin (or vehicle) and at the end of the study (when tumors had developed in most rats). We found an increase in the final body weights, number of mammary cancers, and the final weights of tumors in rats on western diet compared with standard diet. Furthermore, metformin failed to decrease tumor multiplicity in either diet, in fact, it increased final cancer weights (P<.05). Serum from rats in the various groups were examined for approximately 500 metabolites (Metabolon; Research Triangle Park, NC). The Metabolites which were differentially expressed when comparing Western vs standard diets at either time point included myoinositol, scyllo-inositol, 2-hydroxydecanoate, carnitine, tocopherol, and nicotinamide. Certain of these were fatty acids and lipid soluble vitamins that one would expect to be altered by diet. We also compared serum from animals on standard or Western diets at the early or late time points to determine metabolites associated with cancer development. Metabolites whose levels changed in tumor bearing rats included 4-OH-butyrate, acetyl-carnitine, uranate, and threonate. Finally, we compared the effects of metformin treatment in rats on Western or standard diet and found that the metabolic changes associated with metformin treatment were dependent on the diet employed. Thus, metformin altered levels of arachidate and serotonin only in rats on standard diet, while carnnitine and N-delta-acetylornithine were only modulated in rats on Western diet. Again, the most striking result may be that we still failed to observe any efficacy of metformin in animals on standard or Western diet.

Citation Format: Mark S. Miller, Matthew D. Thompson, Ronald A. Lubet, Vernon E. Steele, Harold E. Seifried, Clinton J. Grubbs. Effects of metfornin on mammary carcinogenesis and metabolic profiles in methylnitrosourea-treated rats on standard or Western diets. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 824.
Abstract 827: Polyphenon E treatment affects p73 mRNA levels
Authors: Carastro L. Michael , Cordova Ricardo A. , Cole Christopher D. , Park Jong Y.
Published: July 2016

Prostate cancer (PCa) is the most common malignancy among men in the USA. Polyphenon E (PolyE) is a standardized blend of polyphenols found in green tea extract, which has been shown to have chemoprevention value in PCa models, but the molecular mechanism(s) have not been elucidated. The p73 gene is a member of the p53 tumor suppressor family. A p73 dinucleotide polymorphism (DNP) (rs1801173) is a G4C14-to-A4T14 linked pair of transitions located in exon 2 and lies between the P1 and P2 gene promoters. The P1 and P2 p73 gene promoters are the transcription initiation sites for mRNAs encoding TAp73 and deltaNp73 (DNp73) isoforms, respectively. These p73 mRNA isoforms, TAp73 and áNp73, encode protein isoforms which differ in their N-termini. TAp73 isoforms include the full-length N-terminal sequence and are transcriptionally active. áNp73 isoforms lack the N-terminal trans-activation domain and are dominate negative. Recently, we reported the p73 DNP allele was associated with (1) decreased risk [OR = 0.55, 95%CI = 0.31-0.99] for aggressive PCa and (2) increased TAp73/DNp73 protein isoform ratios in ten human cancer cell lines. We hypothesize that exposure to PolyE induces changes in TAp73/DNp73 mRNA isoform ratios. Our goal for this study is to assess the effect of PolyE on total p73 mRNA levels and TAp73/DNp73 mRNA isoform ratios using two human PCa cell lines with different p73 DNP genotypes: DU145 (wild-type) and PC-3 (heterozygous). Cultured cells were treated with PolyE (100 or 200 mg/L). Cellular RNA was isolated, converted to cDNA and used in TaqMan RT-PCR assays to detect total p73, TAp73 or DNp73 isoform mRNAs. Our data from PolyE treatments of both DU145 and PC-3 cells are consistent with increased p73 mRNA levels (2.05 and 1.60 fold, respectively) and higher TAp73/DNp73 mRNA ratios (1.13 and 1.29 fold, respectively) at the low Poly E level (100 mg/L). At the high PolyE level (200 mg/L), the TAp73/DNp73 mRNA ratios significantly decreased for both DU145 and PC-3 (0.54 and 0.49 respectively). However, the overall p73 mRNA levels was not changed in DU145 (0.91 fold), but increased in PC-3 (1.60 fold) as compared with level in negative control at the high Poly E concentration. Therefore, in this exploratory study, PolyE treatment of the two PCa cell lines tested using a more moderate concentration of 100 mg/L results in higher overall p73 levels and increased TAp73/DNp73 mRNA ratios. This data could provide a potential molecular rationale for the observation that Poly E treatments provide chemopreventative impact against PCa.

Citation Format: L. Michael Carastro, Ricardo A. Cordova, Christopher D. Cole, Jong Y. Park. Polyphenon E treatment affects p73 mRNA levels. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 827.
Abstract 829: Chemoprevention of oxidative stress-associated oral carcinogenesis by sulforaphane depends on NRF2 and the isothiocyanate moiety
Authors: Lan Aixian , Li Wenjun , Liu Yao , Zhang Xinyan , Zhou Shanshan , Palko Olesya , Chen Hao , Kapita Mayanga , Prigge Justin , Schmidt Edward , Chen Xin , Sun Zheng , Chen Xiaoxin Luke
Published: July 2016

Oxidative stress is known to play an important role in oral cancer development. In this study we aimed to examine whether a chemical activator of NRF2, sulforaphane (SFN), may have chemopreventive effects on oxidative stress-associated oral carcinogenesis. We first showed that Nrf2 activation and oxidative damage were commonly seen in human samples of oral leukoplakia. With gene microarray and immunostaining, we found 4-nitroquinoline 1-oxide (4NQO) in drink activated the Nrf2 pathway and produced oxidative damage in mouse tongue. Meanwhile whole exome sequencing of mouse tongue identified mutations consistent with 4NQO's mutagenic profile. Using cultured human oral keratinocytes and 4NQO-treated mouse tongue, we found that SFN pre-treatment activated the NRF2 pathway and inhibited oxidative damage both in vitro and in vivo. On the contrary, a structural analogue of SFN without the isothiocyanate moiety did not have such effects. In a long-term chemoprevention study using wild-type and Nrf2-/- mice, we showed that topical application of SFN activated the NRF2 pathway, inhibited oxidative damage, and prevented 4NQO-induced oral carcinogenesis in an Nrf2-dependent manner. Our data clearly demonstrate that SFN has chemopreventive effects on oxidative stress-associated oral carcinogenesis, and such effects depend on Nrf2 and the isothiocyanate moiety.

Citation Format: Aixian Lan, Wenjun Li, Yao Liu, Xinyan Zhang, Shanshan Zhou, Olesya Palko, Hao Chen, Mayanga Kapita, Justin Prigge, Edward Schmidt, Xin Chen, Zheng Sun, Xiaoxin Luke Chen. Chemoprevention of oxidative stress-associated oral carcinogenesis by sulforaphane depends on NRF2 and the isothiocyanate moiety. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 829.
Abstract 831: c-Myc is a novel target of prostate cancer cell growth inhibition by honokiol
Authors: Singh Krishna B. , Hahm Eun-Ryeong , Singh Shivendra V.
Published: July 2016

Honokiol (HNK) has been shown to inhibit human prostate cancer cell growth in vitro and in vivo, but the underlying mechanism is not fully understood. The present study demonstrates that c-Myc, a key mediator of prostate cancer growth, is a novel target of HNK. Exposure of prostate cancer cells to plasma achievable concentrations of HNK resulted in a marked decrease in levels of total and/or phosphorylated c-Myc. HNK-mediated suppression of c-Myc protein level was due in part to repression of its transcription or protein degradation. We also observed down-regulation of c-Myc in tumors from PC-3 xenografted mice upon oral administration of HNK when compared with control; although the difference was not significant. Stable overexpression of c-Myc conferred protection against HNK-mediated inhibition of colony formation and G0-G1 cell cycle arrest in PC-3 cells. Moreover, HNK suppressed the expression of c-Myc downstream target genes (Cyclin D1 and EZH2) and these effects were restored in Myc-overexpressing PC-3 cells. Consistent with these results, cells with stable knockdown of EZH2 were more sensitive to growth inhibition by HNK compared with control cells. Because androgen receptor (AR) is implicated in regulation of c-Myc expression, we explored the possibility whether AR could affect the expression of c-Myc and its target genes. Indeed, AR overexpression in PC-3 cells enhanced the expression of both c-Myc and its target genes. In conclusion, it is reasonable to postulate that HNK may be effective in retarding growth of c-Myc-driven prostate cancer. This study was supported by the grants CA101753 and CA115498 awarded by the National Cancer Institute.

Citation Format: Krishna B. Singh, Eun-Ryeong Hahm, Shivendra V. Singh. c-Myc is a novel target of prostate cancer cell growth inhibition by honokiol. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 831.
Abstract 839: Novel polypectomy system in a preclinical model of colon cancer
Authors: Ertem Furkan U. , Dashwood Wan-Mohaiza , Rajendran Praveen , Raju Gottumukkala , Rashid Asif , Dashwood Roderick
Published: July 2016

Colonoscopy has been used for over a decade to monitor tumor progression in live animals. However, murine models typically have not involved routine resection of entire lesions, to parallel the clinical scenario in which polyps are observed and removed during a colonoscopy examination. A polyp resection (polypectomy) protocol was developed in the polyposis in rat colon (Pirc) model. Because a major cause of morbidity in such rodent models involves blockage of the colon, polypectomy can be used as a preventive strategy, helping to prolong the lifespan of the animal. Polypectomy in murine models opens the door to longitudinal prevention studies, and determining the molecular changes in resected tumors. A means of specifying the location of each lesion within the colon also was developed in order to track individual polyps in a temporal manner. The “PLC” classification system has three components: P, polyp number from the tail-end; L, location of the polyp from the tail-end; and C, clock face orientation of the polyp within the gut lumen. In summary, we developed a novel murine polypectomy protocol that closely parallels the human clinical scenario during a screening colonoscopy. The procedure employed the Pirc model, but it is directly applicable to longitudinal studies in other murine models of colon cancer. This work was supported by P01 grant CA090890 from the National Cancer Institute, by P30 grant ES023512 from the National Institute of Environmental Health Sciences, by the John S. Dunn Foundation, and by a Chancellor's Research Initiative from Texas A&M University.

Citation Format: Furkan U. Ertem, Wan-Mohaiza Dashwood, Praveen Rajendran, Gottumukkala Raju, Asif Rashid, Roderick Dashwood. Novel polypectomy system in a preclinical model of colon cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 839.