bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4837: Joint effect of arsenic exposure and vascular endothelial growth factor polymorphisms on urothelial carcinoma risk.
Authors: Wang Yuan-Hung , Wu Chia-Chang , Chen Kuan-Chou , Chen Hsin-An , Huang Ming-Te , Wu Chih-Hsiung , Chiou Hung-Yi
Published: August 2014

Arsenic exposure is a major environmental risk factor for Urothelial carcinoma (UC) which includes cancers of bladder, renal pelvis and ureter (upper urinary tract). Vascular endothelial growth factor (VEGF) is an important factor in angiogenesis which plays a critical role in various malignancies including UC. The aim of this study is to investigate the joint effect of arsenic exposure and VEGF polymorphisms on UC risk. A hospital-based case-control study consisted of 730 histologically confirmed UC cases and 850 age-matched cancer-free controls were recruited from September 1998 to December 2009. Polymorphisms in VEGF gene was determined by polymerase chain reaction-restriction fragment length polymorphism. The joint effects on UC risk were estimated by odds ratios (ORs) and 95% confidence intervals (CIs) using unconditional multivariate logistic regression. Subjects with high arsenic exposure had significantly increased risks of 2.4 and 2.5 for bladder cancer and upper urinary tract UC (UUTUC), respectively. A significantly higher UUTUC risk of 1.7 was found in those with one or more risk genotypes of VEGF gene. Individuals with high arsenic exposure who carried one or more risk genotypes of VEGF gene had significantly increased risks of 2.9 (95% CI=1.8-4.7) and 4.2 (95% CI=2.3-7.7) for bladder cancer and UUTUC, respectively. The UUTUC cases with high arsenic exposure carrying one or more risk genotypes of VEGF gene had a higher risk of advanced tumor stage. Our findings suggest that the joint effect of arsenic exposure and risk genotypes of VEGF gene is relatively predominant in UUTUC than in bladder cancer. Keywords: Arsenic, Bladder cancer, Upper urinary tract, Urothelial carcinoma, Vascular endothelial growth factor

Citation Format: Yuan-Hung Wang, Chia-Chang Wu, Kuan-Chou Chen, Hsin-An Chen, Ming-Te Huang, Chih-Hsiung Wu, Hung-Yi Chiou. Joint effect of arsenic exposure and vascular endothelial growth factor polymorphisms on urothelial carcinoma risk. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4837. doi:10.1158/1538-7445.AM2013-4837
Abstract 4838: Exploration of genome-wide two-locus SNP-SNP interactions relate to nasopharyngeal carcinoma susceptibility.
Authors: Su Wen-Hui , Chang Kai-Ping , Shugart Yin Yao , Chang Yu-Sun
Published: August 2014

Most of the genome-wide association studies (GWAS) are concentrated on the detection main effects and can only explain only a small fraction of the genetic risk. One possible explanation might be the existence of interactions between loci. This effect might be missed in traditional GWAS analysis. Testing for SNP-SNP interaction within a single GWAS study can identify the stronger results that are revealed when genes interact. Here, we have completed the genome-wide two-locus SNP-SNP interaction analysis in nasopharyngeal carcinoma (NPC). We started with the NPC GWAS dataset as our Discovery dataset, then we performed an exhaustive genome-wide two-locus search by pairing each SNP sets according to chromosome positions for PLINK epistasis analysis. The analysises were also performed within each SNP sets to ensure every SNP has performed epistasis analysis with all the other SNPs in the dataset. In total, there were 4,244,943 unique interactions after the genome-wide two-locus epistasis analysis. The top 10,000 interaction pairs from the Discovery dataset were selected for validation test in a subset of independent samples. Finally, 67 SNP pairs remain significant in Validation and Combined analysis and can successfully passed permutation tests for 10,000 times. The 67 significant interaction pairs can be clustered into 39 interaction groups which 12 (32%) of the 39 clusters were with at least two nearby SNPs from one of the interaction chromosome. This study represents one of the first attempts to explore the high-dimensional genetic architecture of NPC on a genome-wide scale.

Citation Format: Wen-Hui Su, Kai-Ping Chang, Yin Yao Shugart, Yu-Sun Chang. Exploration of genome-wide two-locus SNP-SNP interactions relate to nasopharyngeal carcinoma susceptibility. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4838. doi:10.1158/1538-7445.AM2013-4838
Abstract 4842: Hazard ratio for colorectal cancer risk in Lynch syndrome is inversely associated with age.
Authors: Jenkins Mark A. , Win Aung Ko , Dowty James G. , Mathews John D. , Hopper John L.
Published: August 2014

Introduction: Carriers of germline mutations in mismatch repair genes (MLH1, MSH2, MSH6, PMS2) have a high risk of colorectal and other cancers (Lynch syndrome). Between 30% and 70% of carriers will be develop colorectal cancer by age 70 if they are not screened for the disease. What is not well understood is how the effect of mutations, in absolute and relative terms, varies by age.

Methods: We conducted a literature review to identify studies of colorectal cancer that estimated age-specific hazard ratios for colorectal cancer for carriers of a mutation in MLH1 or MSH2 compared with the general population. Studies were excluded if they did not adjust correctly for ascertainment. We conducted a random effects meta-analysis to estimate the average age-specific hazard ratio. Annual risk (incidence) by multiplying the population incidence by the age-specific hazard ratio estimated by the meta-analysis.

Results: We identified six studies comprising 673 families (282 MLH1, 391 MSH2). All studies observed that the hazard ratio for colorectal cancer was highest at young ages and decreased with age. This pattern was observed for both males and females and for both MLH1 and MSH2 mutation carriers. Overall, the hazard ratio dropped 6-8 fold over three decades (Table). Incidence of colorectal cancer for carriers increased until age 40 years for males and 50 years for females, and then plateaued (Table).

Conclusion: At younger ages, in multiplicative terms, the magnitude of the increased risk conferred by the mutations is substantially greater than at older ages. The reason for this age-dependency is not known. It is, however, consistent with a multi-stage model of carcinogenesis for Lynch syndrome. These findings add to the complement of factors that complicate risk estimation for Lynch syndrome including: polygenic effects, non-genetic risk modifiers, and the possibility of a parent-of-origin effect. There is still much to learn about the clinical implications and carcinogenesis of Lynch syndrome.

Meta-analysis estimates of hazard ratios and incidences for colorectal cancer by age group. 30-39 year olds 40-49 year olds 50-59 year olds 60-69 year olds Hazard ratio (95%CI): Risk of colorectal cancer in carriers/risk for the general population. Males 120 (60-243) 95 (39-226) 33 (14-78) 14 (4.2-51) Females 80 (42-154) 70 (35-139) 28 (19-41) 14 (7.4-26) Incidence: Proportion of carriers diagnosed with colorectal cancer per year. Males 1 in 146 1 in 50 1 in 39 1 in 38 Females 1 in 490 1 in 135 1 in 69 1 in 61

Citation Format: Mark A. Jenkins, Aung Ko Win, James G. Dowty, John D. Mathews, John L. Hopper. Hazard ratio for colorectal cancer risk in Lynch syndrome is inversely associated with age. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4842. doi:10.1158/1538-7445.AM2013-4842
Abstract 4844: Polymorphisms in regulatory T cell related genes and ovarian cancer survival.
Authors: Charbonneau Bridget , Knutson Keith , Vierkant Robert A. , Cunningham Julie M. , Fogarty Zachary C. , Kalli Kimberly R. , Maurer Matthew J. , Oberg Ann L. , Fridley Brooke L. , Preston Claudia C. , Pharoah Paul D.P. , Ramus Susan J. , Phelan Catherine M. , Odunsi Kunle , Moysich Kirsten , Goode Ellen L.
Published: August 2014

Tumor infiltrating regulatory T cells (Tregs) appear to play a role in survival in a number of solid tumors, including ovarian cancer. We assessed genetic variation via tag SNPs (N=749) in 25 Treg associated genes (CD28, CTLA4, FOXP3, IDO1, IL10, IL10RA, IL15, 1L17RA, IL23A, IL23R, IL2RA, IL6, IL6R, IL8, LGALS1, LGALS9, MAP3K8, STAT5A, STAT5B, TGFB1, TGFB2, TGFB3, TGFBR1, TGRBR2, and TGFBR3) in relation to ovarian cancer survival. SNPs were selected based on r2 ≥ 0.8, MAF ≥ 0.05, 5 kb upstream and downstream from the gene. We had SNP and outcome data for ovarian cancer from 10,084 combined ovarian cancer cases, including 5,248 high grade serous, 1,452 endometrioid, 661 mucinous, and 795 clear cell cases of European descent across multiple studies from the Ovarian Cancer Association Consortium (OCAC). Cox proportional hazards regression modeling was used to estimate per-allele hazard ratios (HRs) and 95% confidence intervals (CIs) for associations with overall survival, adjusting for age at diagnosis, population substructure PCs, study site, histology (for the all case analysis), tumor stage, tumor grade and oral contraceptive use. The strongest associations were found for endometrioid histologic subtype and IL2RA SNPs (maximum r2=0.67) rs11256497 [HR=1.42, 95% CI: 1.22-1.64; p=5.7 x 10−6], rs791587 [HR=1.36, 95% CI: 1.17-1.57; p=6.2 x 10−5], rs2476491 [HR=1.40, 95% CI: 1.19-1.64; p=5.6 x 10−5], and rs10795763 [HR=1.35, 95% CI: 1.17-1.57; p=7.9 x 10−5], as well as clear cell ovarian cancer and CTLA4 SNP rs231775 [HR=0.67, 95% CI: 0.54-0.82; p=9.3 x 10−5]. No other associations were observed at p<1 x 10−4. Overall, this study provided evidence that variation in genes related to Tregs, particularly IL2RA and CTLA4, influence ovarian cancer survival.

Citation Format: Bridget Charbonneau, Keith Knutson, Robert A. Vierkant, Julie M. Cunningham, Zachary C. Fogarty, Kimberly R. Kalli, Matthew J. Maurer, Ann L. Oberg, Brooke L. Fridley, Claudia C. Preston, Paul D.P. Pharoah, Susan J. Ramus, Catherine M. Phelan, Kunle Odunsi, Kirsten Moysich, Ellen L. Goode. Polymorphisms in regulatory T cell related genes and ovarian cancer survival. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4844. doi:10.1158/1538-7445.AM2013-4844
Abstract 4846: Genetic variants in the Wnt/beta-catenin signaling pathway as indicators of ovarian cancer risk and survival.
Authors: Pierzynski Jeanne A. , Hildebrandt Michelle AT , Wu Xifeng , Lu Karen H. , Liang Dong
Published: August 2014

Ovarian cancer is the ninth most common cancer in women and the fifth leading cause of cancer death in 2012. Much is known about the correlation between BRCA mutations and ovarian cancer, but less is known about the impact of common, germline genetic variants on ovarian cancer risk and clinical outcomes. The Wnt/beta-catenin pathway has major roles in regulation of stem cells and other cellular pathways and has been shown to be involved in the development of several different cancers. Therefore, we examined 434 Single Nucleotide Polymorphisms (SNPs) in the Wnt/beta-catenin pathway in 339 ovarian cancer cases and 349 controls. These variants were then assessed for association with risk and clinical outcomes of ovarian cancer. Sixteen polymorphisms had a statistically significant association with ovarian cancer risk with the most significant association being for rs11668593 in APC2 under the dominant model [odds ratio (OR)=0.61; 95% confidence interval (CI), 0.44-0.83]. Unfavorable analysis showed a highly significant cumulative effect of these 16 SNPs on risk (P for trend = 4.22x10−15). In the clinical outcomes analysis, there were eight SNPs that showed an association with ovarian cancer survival. The most significant association with survival was seen for rs11054760 in LRP6 [OR=2.16; 95% CI, 1.33-3.50] which resulted in a poorer survival in patients carrying the heterozygous or homozygous variant genotypes compared to those with the homozygous common genotype. Individuals with the common genotype for rs11054760 had a longer median survival time of 52.7 months compared to only 25.9 months for those with variant containing genotypes (Plog-rank = 3.54x10−4). A gene-dosage effect with a decrease in survival as the number of unfavorable genotypes increased was evident (P for trend = 4.57x10−9). These results implicate genetic variation in the Wnt/beta-catenin signaling pathway as having an impact on risk and clinical outcomes of ovarian cancer.

Citation Format: Jeanne A. Pierzynski, Michelle AT Hildebrandt, Xifeng Wu, Karen H. Lu, Dong Liang. Genetic variants in the Wnt/beta-catenin signaling pathway as indicators of ovarian cancer risk and survival. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4846. doi:10.1158/1538-7445.AM2013-4846
Abstract 4853: Comparison of automated and a semi-automated methods for assessing Ki-67 staining.
Authors: Ondracek Rochelle P. , Marshall James , Kasza Karin , Morrison Christopher , Ghajar Simoya , Morrison Carl , Mehedint Diana , Mohler James L.
Published: August 2014

Human evaluation of immuno-staining is limited by the subjectivity of the human evaluator, is time-consuming and is expensive. Semi-automated means of tissue evaluation have been proposed, but most of these have large subjective components. This study assesses semi-automated and newly-developed fully automated methods of evaluating Ki-67 staining of benign and malignant tissue from radical prostatectomy specimens. A research tissue microarray was constructed from 68 prostatectomy specimens and contained 3 core samples of benign and 3 of malignant tissue from each research subject. Tissue microarray sections were immune-stained for Ki-67. Two technicians (SG & CM) evaluated each TMA core and labeled the nuclei positive for Ki-67 with a green dot, and negative with a red dot. Images were collected and Image-Pro software was used to count the number of green and red dots. In a separate procedure, Image-Pro (IP) software was used to identify nuclei, and grade each nucleus as positive or negative, and the percentage of positive nuclei was calculated.

The semi-automated results correlated with each other (0.58, P<0.001). The semi-automated and fully automated results also correlated (0.8, P<0.001, CM vs. IP and 0.59, P<0.001, SG vs. IP). When stratified by tissue type (benign vs. malignant), the semi-automated results and the fully-automated results also correlated (benign tissue: 0.68, P<0.001, CM vs. IP, and 0.80, P<0.001, SG vs. IP; tumor tissue: 0.89, P<0.001, CM vs. IP and 0.47, P<0.001, SG vs. IP).

These results suggest that semi-automated and automated analysis of Ki-67 staining of prostate tissue produces comparable results, and automated analysis can be used for prostate cancer research.

Citation Format: Rochelle P. Ondracek, James Marshall, Karin Kasza, Christopher Morrison, Simoya Ghajar, Carl Morrison, Diana Mehedint, James L. Mohler. Comparison of automated and a semi-automated methods for assessing Ki-67 staining. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4853. doi:10.1158/1538-7445.AM2013-4853
Abstract 4860: Chemopreventive effects of α-santalol on ultraviolet B induced skin tumorigenesis by modulating cell cycle regulators.
Authors: Santha Sreevidya , Dwivedi Chandradhar
Published: August 2014

Previous studies from our laboratory have shown chemopreventive effects of 5% α-santalol, a sequiterpene isolated from the sandalwood oil in ultraviolet-B induced skin tumor multiplicity in SKH-1 hairless mice. However, this dosage did not have significant effect on tumor incidence. The objective of this study was to investigate the effects of α-santalol (10%, w/v in acetone) on UVB-induced skin tumor incidence in SKH-1 hairless mouse model and to identify the molecular targets in α-santalol mediated protection. Tumor initiation and promotion were carried out by UVB radiation (30 mJ/cm2/day for 5 days a week up to 30 week) that is in the range of human exposure to sunlight that can cause skin cancer. Topical treatment of mice with α-santalol causes a significant reduction in tumor multiplicity, tumor volume and tumor incidence, and also delayed tumor development. Histopathological analysis of hematoxylin-eosin stained skin sections showed that α-santalol pretreatment strongly inhibited UVB-induced epidermal hyperplasia and total thickness of the epidermis. The proliferation potential of skin and tumor tissue sections from UVB-induced carcinogenesis protocol was evaluated by immunohistochemical expression of proliferation marker PCNA. Both intensity of brown staining as well as the number of positively stained nuclei were reduced in skin tumors and adjacent skin tissues from α-santalol treated group when compared to control group. Western blot analysis of skin lysates showed that α-santalol decreases the level of different cyclin-dependent kinases and associated cyclins, Cdc25 phosphatases (Cdc25B and Cdc25C) together with an up-regulation of CDK inhibitor Cip1/p21. However, the level of Kip1/p27 and p53 did not show a significant difference. Our data suggested that α-santalol (10%, topical) is a safer and promising skin cancer chemopreventive agent through targeting different cell cycle regulators. (This work is supported by a Translational Cancer Center Research Grant, funded by the State of South Dakota).

Citation Format: Sreevidya Santha, Chandradhar Dwivedi. Chemopreventive effects of α-santalol on ultraviolet B induced skin tumorigenesis by modulating cell cycle regulators. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4860. doi:10.1158/1538-7445.AM2013-4860
Abstract 4862: Cancer chemopreventive potential of grape seeds and skin in mice inoculated with Ehrlich ascites carcinoma cells.
Authors: Badr El-Din Nariman K. , Ali Doaa A. , Abou-El-Magd Rania F. , Soliman Nahed A.
Published: August 2014

Chemoprevention by dietary constituents holds great promise to control cancer. Grape seeds(GSE) and skin (GSK) contain large amounts of phytochemicals such as flavonoids, phenolic acids, and anthocyanidins, which play an important role as chemopreventive and anticancer agents. This study was undertaken to investigate the chemopreventive effects of GSE and GSK efficacies and associated mechanisms on tumor growth in mice. GSE & GSK were separated from the pulp of grape Vitis vinifera, dried and ground to powder. Equal amounts of GSE & GSK were mixed uniformly with the standard diet powder at concentrations of 10% (w/w). The supplemented diet was started two weeks before tumor cells inoculation and continued until the end of the experiment. Female Swiss albino mice were inoculated subcutaneously with 2.5x106

Ehrlich ascites carcinoma (EAC) cells. Tumor growth was monitored for 30 days.Cell cycle progression was measured and the apoptotic effect of GSE and GSK on tumor cells were evaluated by flow cytometric analysis, histopathology and transmission electron microscopy examination. GSE & GSK intake prevented tumor development in 47% of the animals.

Tumor volume and tumor weight measured at the termination of the experiment were reduced by 93.9 %& 86.3 %, p< 0.001) respectively as compared to the untreated tumor- bearing mice. GSE& GSK induced a marked increase in the percentage of apoptotic tumor cells as determined by flow cytometric analysis and confirmed by histopathological and electron microscopy examination. These changes were associated with significant

G1-phase cell cycle arrest, decrease in the expression of Bcl-2, increase of the expression of p53 and induced caspase-3 activation in the tumor cells. Our data demonstrated that grape seeds& skin intake resulted in profound cell growth inhibition by a mechanism involved G1-phase cell cycle arrest, and apoptosis. Therefore, dietary grape seeds and grape skin possess cancer chemopreventive potential and provide support for their use in human cancer chemoprevention.

Citation Format: Nariman K. Badr El-Din, Doaa A. Ali, Rania F. Abou-El-Magd, Nahed A. Soliman. Cancer chemopreventive potential of grape seeds and skin in mice inoculated with Ehrlich ascites carcinoma cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4862. doi:10.1158/1538-7445.AM2013-4862
Abstract 4864: Aroma terpenes prevent UVB-induced sunburn in Skh1 mouse skin.
Authors: Uddin Ahmed N. , Labuda Ivica , Wu Feng , Tchou-Wong KamMeng , Burns Fredric J.
Published: August 2014

Understanding how aroma terpenes prevent sunburn and/or skin cancer in mice could lead to more effective and safer ways of blocking sun damage to human skin. In our previous study, β-damascenone, an aroma terpene, protected against sunburn by activating both keratinocyte and sebaceous gland pathways that fortified and thickened the cornified envelope and sebum layers of mouse epidermis. In the current study, the sunburn preventive activity of d-limonene was examined to confirm that these related compounds work similarly. In addition to sunburn, immunohistochemical analyses of proliferation (PCNA) and a damage-response gene N-Myc Down Regulated Gene 1 (NDRG1) were investigated in control versus d-limonene-treated Skh-1 mice. The mice were treated either topically on the dorsum or orally with different concentrations of d-limonene in liponate (100%, 10%, 1% and 0%) followed by exposure of the dorsum to solar simulation ultraviolet radiation (UVR) of 1.5 kJ/m2 from fluorescent ‘sunlamps’. Either 4 topical 5 μL doses or a single dose of 20 μL (0.95 μg/g body weight) of 100% d-limonene provided complete protection from UVR-induced sunburn. The protection was less but still significant at 10% d-limonene but was absent at 1% d-limonene in comparison to UVR alone. PCNA-labeling showed a marked proliferation increase of epidermal basal cells, outer root sheath cells of hair follicles and sebaceous gland cells by separate exposures to d-limonene and UVR relative to controls. The combined treatment of d-limonene followed by UVR produced fewer PCNA-positive cells in epithelial tissue relative to UVR-only mice. The NDRG1 protein (an indirect measure of DNA damage) was overwhelmingly (∼85% of keratinocytes) induced by UVR indicating heavy DNA damage. The UVR-induced NDRG1 index remained at ∼85% at 1% d -limonene, but was significantly reduced at 10% d-limonene to ∼24% and to less than 1% at 100% d -limonene indicating nearly complete elimination of cellular damage as a pure compound; a finding that is consistent with complete sunburn prevention. The thickness of the cornified envelope plus sebum layers nearly doubled within 5 days after administration of the d-limonene corresponding to the time of UVR application. Overall the results indicate that d-limonene protected against sunburn by activating keratinocyte/sebaceous gland-based pathways that fortified and thickened the cornified envelope plus sebum layers on the skin surface by releasing elevated levels of UVR-absorbing proteins that decreased UVR dose to underlying cutaneous tissues.

Citation Format: Ahmed N. Uddin, Ivica Labuda, Feng Wu, KamMeng Tchou-Wong, Fredric J. Burns. Aroma terpenes prevent UVB-induced sunburn in Skh1 mouse skin. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4864. doi:10.1158/1538-7445.AM2013-4864
Abstract 4867: The metabolomic signature of Rhodiola rosea L. extracts- (SHR-5) treated mouse bladder cancer in the UPII-mutant Ha-ras transgenic model.
Authors: Liu Zhongbo , Blair Christopher A. , Zi Xiaolin
Published: August 2014

Rhodiola rosea has been used for centuries in the traditional medicine for reducing depression, enhancing work performance, and preventing high altitude sickness. We have recently shown that Rhodiola rosea L. extract, SHR-5, markedly inhibited the growth of bladder cancer and the survival of mice in the UP-II-mutant Ha-ras transgenic model via inhibition of the mTOR pathway. The purpose of this study was to further investigate the effects of SHR-5 on bladder tumor metabolism. The global metabolic profiles were compared for bladder tumors from mice that were fed with normal drinking water (n=7), 1.25 (n=7) or 6.25 (n=7) mg/ml SHR-5 in water for about five months. The analysis was conducted at Metabolon Inc. with a combination of high-throughput LC- and GC-based MS method. From a metabolomics library consisting of more than 2,000 standards, a total of 341 named metabolites were detected. Phenolic metabolites, such as phenylpropionylglycine, hydroxycinnamate and catechol sulfate, were increased, while prostaglandin metabolites, such as prostaglandins A2, D2 and E2, were decreased in tumors treated with SHR-5. This observation suggested the potential anti-inflammation effect of SHR-5 in bladder tumors. Neurotransmitters GABA, taurine, NAA and 2-oxoindole-3-acetate was more abundant; whereas kynurenine, histamine and aminoadipate were less abundant with SHR-5 treatment. This result is consistent with mood-improving effects of SHR-5. In addition, SHR-5 treatment resulted in increased levels of glucose and glycogen metabolites (e.g. maltotriose, maltose), as well as two glycolysis intermediates (3-phosphoglycerate and 2-phosphoglycerate) and the citric acid cycle (TCA) intermediates (fumarate and malate) compared to control treatment. These patterns suggest that SHR-5 may increase mobilization of glycogen for energy production in glycolysis and TCA cycle. These effects of SHR-5 may be mediated by differential effects on the mTOR pathway. Taken together, we show that global metabolomic profiling provides a unique and efficient tool for studying mechanisms of complex herb extracts’ action.

Citation Format: Zhongbo Liu, Christopher A. Blair, Xiaolin Zi. The metabolomic signature of Rhodiola rosea L. extracts- (SHR-5) treated mouse bladder cancer in the UPII-mutant Ha-ras transgenic model. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4867. doi:10.1158/1538-7445.AM2013-4867
Abstract 4873: Genistein and soy isoflavone isolate decreased tumor incidence and multiplicity in a Sprague-Dawley ER-positive breast cancer model.
Authors: Nicastro Holly L. , Lubet Ronald A. , Dunn Barbara K. , Grubbs Clinton J.
Published: August 2014

This study evaluated the chemopreventive efficacy of the pure soy isoflavone genistein and of a soy isoflavone mixture (PTI G-4660) containing roughly 90% genistein in an ER-positive breast cancer model in rats. Female Sprague Dawley rats were injected i.v. with N-nitroso-N -methylurea (MNU) at 50 days of age. In a model examining the effects on cancer initiation, promotion, and progression, rats were fed either a standard Teklad diet or a diet supplemented with genistein (200 or 2000 mg/kg diet) beginning at 43 d of age. Supplemented or standard diets were administered continuously thereafter until the end of the study. In a separate study testing the effects on progression, rats were administered standard Teklad diet or diet supplemented with PTI G-4660 (2000 mg/kg diet) beginning at 55 d of age. All rats were weighed weekly, palpated for mammary tumors twice per week, and checked daily for signs of toxicity. The studies were terminated 126 d after MNU treatment and tumors were weighed. After 126 days, genistein at 200 and 2000 mg/kg diet decreased tumor incidence in a dose-dependent fashion by 15 and 20% (p<0.05), respectively, and significantly decreased tumor multiplicity by 49.6 and 60.3% (p<0.05), respectively. PTI non-significantly decreased tumor incidence by 14% and tumor multiplicity by 24%. In addition, gene expression profiling of livers from rats treated with standard Teklad diet or genistein revealed pathways modified by genistein which may be useful as biomarkers.

Citation Format: Holly L. Nicastro, Ronald A. Lubet, Barbara K. Dunn, Clinton J. Grubbs. Genistein and soy isoflavone isolate decreased tumor incidence and multiplicity in a Sprague-Dawley ER-positive breast cancer model. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4873. doi:10.1158/1538-7445.AM2013-4873
Abstract 4874: Bisphenol A exposure negates tamoxifen-mediated chemoprevention in MMTV-erbB2 transgenic mice.
Authors: Yang XiaoHe , Ma Zhikun , Jiang Yunbo
Published: August 2014

Bisphenol A (BPA), the building block of polycarbonate plastics with estrogenic activity, is one of the most common chemicals people are exposed to in daily life. Increasing reports suggest a link between BPA exposure and increased breast cancer risk. Although the effect of early BPA exposure and breast cancer risk later in life has been extensively studied, whether this xenoestrogen interferes with hormonal therapeutic agents, such as tamoxifen, has hardly been documented. In this study, we tested the effect of BPA exposure on mammary tumor development and tamoxifen-mediated chemoprevention in MMTV-erbB2 transgenic mice. BPA (500 ng/kg in sesame oil) was administered daily by subcutaneous injection for 8 weeks, starting from 8 weeks of age. Citrate tamoxifen at a dose of 1 mg/kg/day was administered by oral gavage alone or in combination with BPA. Based on tumor latency, we found that BPA alone accelerated mammary tumor development whereas tamoxifen treatment profoundly delayed the tumorigenesis. A combination of tamoxifen-treatment and BPA exposure, however, significantly (p<0.01) interfered with tamoxifen-mediated prevention as indicated by a shortened tumor latency compared to the tamoxifen group. BrdU incorporation and whole mount analysis of mammary glands at 16 weeks showed that BPA or tamoxifen alone induced distinctive proliferative and inhibitory ductal growth respectively. A combination of tamoxifen and BPA exposure abrogated tamoxifen-mediated growth inhibition. Importantly, altered tumor development and morphogenesis in each group were accompanied by modified signaling in estrogen receptor (ER) and EGFR/erbB2 pathways in mammary tissues. BPA alone induced enhanced phosphorylation/activation of ERα, the receptor tyrosine kinases (erbB2, EGFR and erbB3) and downstream targets (AKT1, and ERK½). Protein and mRNA levels of ERα, cyclin D1, EGFR, erbB3, and c-myc were also increased. In contrast, tamoxifen inhibited the activation of both pathways, as indicated by decreased phosphorylation and expression of those markers. In the combination group BPA exposure blocked tamoxifen-mediated inhibition of ER and erbB2 signaling. Our results demonstrated that BPA exposure promotes erbB2-associated carcinogenesis and negates tamoxifen-mediated chemoprevention. The underlying mechanisms involve BPA-induced activation and crosstalk of ER and erbB2 pathways. Taken together, our data suggests that BPA exposure is a risk factor that may interfere with hormonal therapy, which is of great translational value.

Citation Format: XiaoHe Yang, Zhikun Ma, Yunbo Jiang. Bisphenol A exposure negates tamoxifen-mediated chemoprevention in MMTV-erbB2 transgenic mice. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4874. doi:10.1158/1538-7445.AM2013-4874
Abstract 4881: Influence of polyphenols on selenoprotein expression in human colon cancer cells.
Authors: Galinn Sarah E. , Rosenthal Lindsay C. , Barsotti Steven M. , Tsuji Petra A.
Published: August 2014

Selenium, an essential micronutrient, is recognized as a cancer protective agent that functions through low-molecular weight selenocompounds, and, importantly, through selenoproteins. Previous studies suggest that the 15 kDa selenoprotein (Sep15), a gene polymorphic in humans, has an important role in colon cancer in vivo, and that reduction of Sep15 in human HCT116 colon cancer cells reverses their cancer phenotype. Our diet contains a wide array of nutrients, and it is conceivable that these share molecular targets with selenium and regulate selenoprotein expression. The selenoproteins Thioredoxin Reductase 1 (TR1) and Glutathione Peroxidase (GPx) 2 are regulated by the transcription factor Nrf2, whereas GPx1 is not. Some dietary compounds, e.g., Sulforaphane, are known Nrf2-inducers, impacting expression of TR1 and GPx2. However, the effect of Sulforaphane and many polyphenols on other selenoproteins, or, how other polyphenols influence TR1 and GPx2 remains to be elucidated. Using HCT116 cells, we are investigating the effects of polyphenols and Sulforaphane on selenoprotein expression. Cells with wild type or down-regulated expression of Sep15 were exposed to 5 μM Sulforaphane or 20 μM 5,7-Dimethoxyflavone (5,7-DMF) for 48 hours. Subsequently, mRNA was isolated and examined with real-time RT-PCR. Our preliminary results suggest that, Sulforaphane exposure resulted in a slight increase of TR1 mRNA in both control and shSep15 cells, but not GPx2. Interestingly, 5,7-DMF exposure resulted in increased GPx1 (P<0.01) and GPx2 (P>0.05) mRNA in control cells. Surprisingly, GPx2 mRNA appeared significantly down-regulated (P<0.01) in shSep15 cells compared to DMSO treated cells, whereas GPx1 mRNA appeared unaffected. Western blotting and activity measurements are currently underway to support these results.

Citation Format: Sarah E. Galinn, Lindsay C. Rosenthal, Steven M. Barsotti, Petra A. Tsuji. Influence of polyphenols on selenoprotein expression in human colon cancer cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4881. doi:10.1158/1538-7445.AM2013-4881
Abstract 4884: Involvement of autophagy in penta-1,2,3,4,6-O-galloyl-beta-D-glucose-induced senescence-like growth arrest in human cancer cells.
Authors: Dong Yinhui , Lü Junxuan , Hu Hongbo
Published: August 2014

Autophagy and senescence are two important cellular processes that have been shown to play critical roles in the regulation of carcinogenesis. Our previous studies have demonstrated that cellular features of both autophagy and senescence were induced in response to penta-1,2,3,4,6-O-galloyl-beta-D-glucose (PGG), a polyphenolic anti-cancer compound, in multiple types of cancer cells. In this follow-up study, we further investigated the cross-talk among these PGG-induced phenotypes and signal transduction mechanisms. In cell culture models, we show that inhibition of autophagy by either chemical inhibitors or genetic approaches (RNA interference) led to a near abolishment of PGG-induced senescence, followed by induction of apoptosis, indicating autophagy is required for the senescent phenotype induced by PGG and antagonistic of apoptosis execution. In addition, we identify unfolded protein responses (UPR) in the endoplasmic reticulum (ER) to play a pivotal role in autophagy-mediated senescence phenotype. Furthermore, our data show that c-Jun NH(2)-terminal kinase (JNK) could be a critical mediator for PGG-induced autophagy. In a subcutaneous xenograft mouse model of human HepG2 liver cancer, we detected biochemical markers of modulations of autophagy, ER stress and senescence induced by PGG intra-peritoneal administration for 16 days. Our findings provided novel insights into cellular processes and signaling mechanisms involved in PGG-induced senescence-like growth arrest in human cancer cells.

Citation Format: Yinhui Dong, Junxuan Lü, Hongbo Hu. Involvement of autophagy in penta-1,2,3,4,6-O-galloyl-beta-D-glucose-induced senescence-like growth arrest in human cancer cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4884. doi:10.1158/1538-7445.AM2013-4884
Abstract 4887: FTY720 prevent tumorigenesis and suppress the progression of colitis-associated cancer.
Authors: Yamada Akimitsu , Liang Jie , Nagahashi Masayuki , Kim Eugene Y. , Harikumar Kuzhuvelil B. , Huang Wei-Ching , Hait Nitai C. , Allegood Jeremy C. , Price Megan M. , Avni Dorit , Kordula Tomasz , Sheldon Milstien , Takabe Kazuaki , Spiegel Sarah
Published: August 2014

Inflammatory bowel disease is an important risk factor for colorectal cancer. Sphingosine-1-phosphate (S1P), a pleiotropic bioactive sphingolipid metabolite formed by two sphingosine kinases, SphK1 and SphK2, is involved in inflammation and cancer. Enhanced tumor number and size, with a concomitant increase in tumor load was observed in mice overexpressing SphK1 treated with the colonotropic mutagen azoxymethane (AOM) followed by three rounds of oral administration of the luminal toxin dextran sodium sulfate (DSS) to induce colitis-associated cancer (CAC). There was also a large increase in adenomas with a high grade of dysplasia associated with increased SphK1 expression and formation of S1P. We demonstrated that the SphK1/S1P axis is important for production of IL-6 and is involved in persistent activation of STAT3 and consequent upregulation of one of its target genes, the S1P receptor S1PR1. Because the pro-drug FTY720 modulates the immune system by acting as a functional antagonist of S1PR1, inducing its internalization and degradation, we examined its effect on CAC progression. FTY720 drastically reduced colitis-associated tumorigenesis, when it was administered during CAC induction. Remarkably, FTY720 blocked CAC progression even when it was administered later after the last DSS treatment when tumors were already established. Moreover, it reduced SphK1 expression, production of IL-6, and persistent activation of STAT3. These findings suggest that FTY720 may be beneficial for preventing tumor formation in individuals with inflammatory bowel disease.

This work was supported by grants from the National Institute of Health (R37GM043880, R01CA61774, U19AI077435 to S.S. and R01CA160688 to K.T.). M.N. is a Japan Society for the Promotion of Science Postdoctoral Fellow.

Citation Format: Akimitsu Yamada, Jie Liang, Masayuki Nagahashi, Eugene Y. Kim, Kuzhuvelil B. Harikumar, Wei-Ching Huang, Nitai C. Hait, Jeremy C. Allegood, Megan M. Price, Dorit Avni, Tomasz Kordula, Milstien Sheldon, Kazuaki Takabe, Sarah Spiegel. FTY720 prevent tumorigenesis and suppress the progression of colitis-associated cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4887. doi:10.1158/1538-7445.AM2013-4887
Abstract 4888: Axl receptor signaling in required for stem cell traits and metastasis in breast cancer.
Authors: Tiron Crina , Pelissier Fanny , Wnuk-Lipinska Katarzyna , Stefansson Ingunn , Virtakoivu Reeta , Miyano Masaru , Sandal Tone , Micklem David , Garbe James , Stampfer Martha , Ivaska Johanna , Akslen Lars , LaBarge Mark , Lorens James
Published: August 2014

Epithelial-to-mesenchymal transition (EMT) endows carcinoma cells with migratory, survival and stem cell-like attributes that facilitate therapeutic resistance and metastasis. Expression of the Axl receptor tyrosine kinase in aggressive breast cancer correlates with EMT, poor survival and is prevalent in patient metastases. Induction of EMT in immortalized mammary epithelial cells by EMT-transcription factors, TGFbeta or hypoxia upregulates Axl and establishes an autocrine-signaling loop with its ligand, Gas6. Axl receptor signaling is required to maintain EMT-related invasive, drug resistance and cancer stem cell (CSC) traits of malignant breast cancer cells. Targeting Axl signaling with RNAi or pharmacological agents blocks the EMT/CSC gene program and inhibits malignant functions including invasiveness, drug resistance, mammosphere formation, in vivo tumor initiation, and spontaneous metastasis in orthotopic breast cancer models. These results suggest that Axl expression is requisite for the maintenance of EMT and stem cell-like traits during malignant progression. The EMT program is characteristic of normal adult mammary epithelial stem/progenitor cells. Congruently, we show that the Axl receptor expressed on mammary epithelial stem/progenitor cells, and Axl signaling is necessary for mammary epithelial multipotent progenitor activity. Thus Axl receptor signaling represents a novel regulatory pathway linking normal mammary stem/progenitor cells and breast cancer stem cells. Hence, clinical Axl inhibitors represent a novel therapeutic avenue to target EMT/CSC traits of aggressive breast cancer.

Citation Format: Crina Tiron, Fanny Pelissier, Katarzyna Wnuk-Lipinska, Ingunn Stefansson, Reeta Virtakoivu, Masaru Miyano, Tone Sandal, David Micklem, James Garbe, Martha Stampfer, Johanna Ivaska, Lars Akslen, Mark LaBarge, James Lorens. Axl receptor signaling in required for stem cell traits and metastasis in breast cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4888. doi:10.1158/1538-7445.AM2013-4888
Abstract 4890: Periostin signaling regulates breast cancer stem cells.
Authors: Lambert Arthur W. , Ozturk Sait , Papageorgis Panos , Abdolmaleky Hamid , Thiagalingam Sam
Published: August 2014

Recent evidence suggests that the molecular heterogeneity inherent to breast cancer, which underlies metastasis, resistance to treatment and disease recurrence, can be driven by a distinct subpopulation of tumor cells referred to as cancer stem cells (CSCs). However, the extracellular cues and intracellular pathways that CSCs rely on remain unclear. Working with a breast cancer progression model system we found the invasive breast cancer cell line to exhibit numerous stem-like characteristics. Gene expression profiling, along with a careful review of the literature, identified a secreted extracellular matrix molecule, periostin (POSTN), which we hypothesized to be required for the maintenance of a cancer stem cell state. POSTN expression was increased in numerous basal-like breast cancer cell lines that contain an expanded CSC population, as well as in cells grown as mammospheres and in the CD44+/CD24- fraction of tumor cells. Furthermore, CSC-like lines displayed increased surface levels of the alpha-v beta-3 integrin heterodimer, a known receptor for POSTN, suggesting that hyperactivation of a POSTN signaling axis could potentially regulate the CSC state. Using stable cell lines expressing an shRNA directed at POSTN we have found that POSTN is required for efficient mammosphere formation and for the maintenance of an ALDH-positive CSC population. In line with this, we found these cells had impaired Erk and Wnt signaling and expressed less IL-6 and IL-8, known regulators of CSCs. Furthermore, high POSTN expression correlates with a worse clinical prognosis in basal-like breast cancer patients. Therefore, we suggest that POSTN signaling through integrin receptors may enrich for highly tumorigenic breast CSCs in a subset of basal-like tumors, leading to aggressive and recurrent malignancies. More broadly, our results imply that a supportive tumor niche can be established and maintained by cancer cells, and raises the possibility of targeting components of this extracellular environment to eradicate breast CSCs.

Citation Format: Arthur W. Lambert, Sait Ozturk, Panos Papageorgis, Hamid Abdolmaleky, Sam Thiagalingam. Periostin signaling regulates breast cancer stem cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4890. doi:10.1158/1538-7445.AM2013-4890
Abstract 4894: Sorafenib enriches for sarcoma cancer stem cells in vitro.
Authors: Canter Robert J. , Ames Erik , Tellez Joseph , Smith Rachel C. , Murphy William J.
Published: August 2014

Background: Cancer stem cells (CSCs) are a putative source of relapse in many cancers following anti-proliferative therapies. We hypothesized that tyrosine kinase inhibition (TKI) and radiotherapy (RT) would enrich for CSCs in sarcoma cell lines.

Methods: A673 Ewing's sarcoma and SW982 synovial sarcoma cells were exposed to sorafenib and RT in a dose- and time-dependent fashion. Cultured cells were harvested and stained with fluorochrome-conjugated antibodies against human CSC markers including CD133, CD24, CD44, and aldehyde dehydrogenase (ALDH). Cell viability was analyzed using 7-Aminoactinomycin (7-AAD). Data were acquired using a BD Fortessa cell sorter (BD Biosciences) and analyzed with FlowJo software version 7.2. Parametric and non-parametric statistical tests were performed as appropriate.

Results: Short-term exposure (≤1 day) to sorafenib demonstrated a linear dose-dependent cytotoxicity for A673 cells at doses ≥ 4 μM, while SW982 cells required doses ≥ 16 μM. At 24 hours, A673 cell viability decreased from 100% at 1 μM sorafenib to 0% at 32 μM (P<0.05). By day 5, A673 cells exposed to sorafenib doses < 32 μM recovered log-phase growth, while cells exposed to doses > 32 μM remained non-viable. Baseline CSC phenotyping of A673 cells demonstrated 55±5% CD133+, 12±7% ALDH+, 14% CD44+, and 0% CD24+ sub-populations. Baseline CSC phenotyping of SW982 cells demonstrated 0.2±0.9% CD133+, 42±8% ALDH+, 95±5% CD44+, and 40±7% CD24+. A673 cells exposed to 24h sorafenib increased the ALDH+ fraction to 40±3% (> 3-fold increase) at sorafenib 16 μM (P<0.05). SW982 cells showed a non-significant increase in the ALDH+ fraction at doses ≤ 16 μM. Although single fractions of RT were anti-proliferative to A673 cells starting at doses ≥5 Gy, there was not an additive effect of 2.5 Gy RT with sorafenib on cell viability or CSC marker expression.

Conclusion: Sorafenib exerts an anti-proliferative effect on sarcoma cells but enriches for sarcoma CSC. The magnitude of these effects appears to be inversely correlated to baseline ALDH+ levels. Sustained anti-sarcoma therapeutic effects may require targeting of the CSC population following anti-proliferative therapy.

Citation Format: Robert J. Canter, Erik Ames, Joseph Tellez, Rachel C. Smith, William J. Murphy. Sorafenib enriches for sarcoma cancer stem cells in vitro. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4894. doi:10.1158/1538-7445.AM2013-4894

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.