bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4594: Grape seed extract induces apoptosis in human colorectal carcinoma through modulation of death receptors and anti-apoptotic molecules
Authors: Derry Molly M. , Agarwal Rajesh , Agarwal Chapla
Published: June 2012

Colorectal Cancer (CRC) is the second leading cause of cancer related deaths in the US. Diets rich in fruits and vegetables are shown to decrease CRC incidence by 40%, in part due to antioxidant and free radical properties of the polyphenolics present therein; grape seed extract (GSE) is one of the richest sources of these antioxidants. Oncogenic mutations that suppress apoptosis can lead to tumor initiation, progression, and metastasis; a major cause of conventional treatment resistance is due to apoptotic pathway defects. Accordingly, the strategies that target the mechanisms of apoptosis resistance in cancer cells provide the potential opportunities for cancer management. Herein we investigated the apoptotic effects of GSE and defined associated mechanisms in three human CRC cell lines, namely SW480, SW620 and HCT116, representing II-IV stages of CRC. Normal colon epithelial cells namely NCM460 was also included to establish GSE specificity, if any, towards CRC cells. A comparison of GSE activity in three CRC cell lines showed that almost three times higher GSE levels are needed for comparable cell death effect in SW480 versus SW620/HCT116 cells; cell treatments were accordingly adjusted for remainder studies. GSE treatment of SW480 cells at 25-100μg/mL resulted in a maximum of 55% apoptotic death at 48h; however, even 10-30μg/mL GSE treatments produced 53-57% apoptotic cell death in SW620 and HCT116 cell lines. At the molecular level, GSE induced DR4/5 up regulation, caspase-9, -3, -8, and PARP cleavage in all three cell lines indicating the involvement of both extrinsic and intrinsic apoptotic pathways in its efficacy. Furthermore, GSE treatment caused down regulation of anti-apoptotic proteins Bcl-2 and XIAP in SW480 cells only, but increased the levels of pro-apoptotic proteins such as Mcl-1s in all three cell lines. Importantly, GSE also increased the levels of p53 and its associated regulators namely p21, PUMA and Bak, suggesting their possible role in GSE-caused apoptotic death in HCT116 cell line. GSE treatment (25-100μg/mL) of NCM460 cells did not result in significant death compared to what was observed in three different human CRC cell lines, clearly showing its activity selectively in cancer cells. In conclusion, our findings further support the notion that GSE could be an effective chemopreventive and chemotherapeutic agent for colorectal.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4594. doi:10.1158/1538-7445.AM2011-4594
Abstract 4599: Apigenin intake suppresses IGF-I signaling in an autochthonous mouse model of prostate cancer
Authors: Shukla Sanjeev , Gupta Sanjay
Published: June 2012

Insulin-like growth factor-I (IGF-I) and its receptor (IR) plays an important role in prostate cancer development and progression. IGF-I binds to the IGF-IR that transduces signals to the nucleus and mitochondria primarily via the mitogen-activated protein kinase (MAPK) and PI3K/Akt pathways facilitating increased cell survival and proliferation. Agents that are capable of inhibiting IGF-signaling pathway might be useful in the prevention and/or therapy of prostate cancer. Apigenin, a common dietary flavonoid abundantly present in fruits and vegetables has shown remarkable cancer preventive and cancer therapeutic effects. The mechanisms underlying the cancer-protective effect remains elusive. We determined the role of IGF-signaling and its downstream and other associated events by apigenin using TRAMP model. TRAMP represents males spontaneously develop prostatic adenocarcinoma that progresses through multiple stages and exhibits both histological and molecular features similar to that of human prostate cancer. TRAMP males exhibit an increase in the level of serum IGF-I, uPA and VEGF; p-Akt (Ser473), p-ERK1 (T202/Y204) and p-ERK2 (T185/Y187) in the dorso-lateral prostate of TRAMP mice in age-dependent manner, compared to non-transgenic littermates. Oral intake of apigenin at 20- and 50- microg/mouse/day (gavaged in 0.2 ml vehicle containing 0.5% methyl cellulose and 0.025% Tween 20) was provided for six days per week for 20 weeks; whereas the third group received vehicle only and served as control. Apigenin intake by TRAMP mice from 8-28 weeks of age exhibited marked reduction in tumor growth with no signs of metastasis and also resulted in a significant decrease in the wet weight of dorso-lateral and ventral lobes of the prostate and genitourinary apparatus. Apigenin intake exhibited a marked dose-dependent reduction in the levels of IGF-I along with significant increase in the levels of IGFBP-3 in the serum of TRAMP mice. The modulation of IGF-I/IGFBP3 was associated with suppression of p-ERK1 (T202/Y204) and p-ERK2 (T185/Y187) and p-Akt (Ser473) levels measured in the tissue lysates from the dorso-lateral prostates of these mice. Furthermore, apigenin intake resulted in marked inhibition of the target genes regulating angiogenesis and metastasis, notably VEGF, uPA, MMP-2 and -9 in the mouse serum. Taken together, this study provides evidence that in vivo growth inhibitory effects of apigenin involve inhibition of IGF-I signaling axis in prostate cancer and further support the development of apigenin as a promising anticancer agent.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4599. doi:10.1158/1538-7445.AM2011-4599
Abstract 46: COSMIC: Enhancing the analysis of whole somatic mutant tumor genomes
Authors: Forbes Simon A. , Bindal Nidhi , Kok Chai Yin , Jia Mingming , Leung Kenric , Gunasekaran Prasad , Bamford Sally , Cole Charlotte , Teague Jon , Stratton Michael R. , Campbell Peter , Futreal P Andrew
Published: June 2012

COSMIC, the Catalogue Of Somatic Mutations In Cancer (http://www.sanger.ac.uk/cosmic), is a comprehensive web resource for the investigation of somatic mutations in human cancer with data curated and centrally combined from a variety of sources. Currently, point mutation data have been manually curated from the scientific literature on 89 genes, together with full mutation details on a further 57 fusion gene pairs (Release v50, Nov 2010), defined as known cancer genes in the Cancer Gene Census (http://www.sanger.ac.uk/genetics/CGP/Census). Further, in collaboration with the IARC TP53 database, COSMIC now contains the somatic mutation data for the key p53 tumor suppressor gene. Details of many large candidate gene screens, together spanning almost 20,000 genes have also been curated from three sources, the scientific literature, the Cancer Genome Project labs at the Sanger Institute, UK (CGP), and The Cancer Genome Atlas (TCGA) web portal. Finally, COSMIC now presents the results of 48 whole-genome screens, detailing 2689 genomic rearrangements, 1661 coding mutations and 55,440 non-coding mutations.

The COSMIC website is designed to allow easy and intuitive browsing of this data, in isolation or various combinations. As the amount and variability of data has increased, the methods of acquiring and interacting with the data have been enhanced. The COSMIC website now provides mining capabilities, with numerous specialization filters, graphically presenting selected data. A more extensive mining tool is the COSMIC Biomart, allowing selection of data using any definitions found in the COSMIC system.

With the increasing significance of genomic data in cancer, COSMIC has integrated with the two largest genome browsers, the Ensembl system and the UCSC cancer genomics browser; links are available from and to Ensembl, whilst per-tissue summaries are provided graphically in the UCSC browser. In addition, COSMIC has integrated its own genome browser, a version of ‘Gbrowse’, which provides custom genomic views of the majority of COSMIC data, together with CONAN copy number graphs, within the context of full genome annotations. Looking to the future, much novel global cancer genomics data will soon be made available via the ICGC data coordination centre. COSMIC is well placed to utilize this as a major new source of mutation information, maintaining COSMIC's position as a central resource in human cancer genetics.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 46. doi:10.1158/1538-7445.AM2011-46
Abstract 4602: Cholesterol and ezetimibe: Effects on tumor growth and survival outcomes: An interim analysis
Authors: Chu David I. , Masko Elizabeth M. , Poulton Susan H. , Pizzo Salvatore V. , Freedland Stephen J.
Published: June 2012

Introduction and Objectives: Prostate cancer (PC) is affected by cholesterol metabolism and cholesterol-mediating medications. Ezetimibe, a gut cholesterol-uptake inhibitor, inhibits angiogenesis in human PC xenografts when given as preventive therapy. We investigated the effect of ezetimibe on PC growth when given as treatment therapy.

Methods: We randomized 125 SCID mice into three initial diets: a high-fat, low-cholesterol diet (HFLC) (40% fat, 43% carbohydrates, 17% protein, 0.21% cholesterol), a high-fat, high-cholesterol diet (HFHC) (40% fat, 43% carbohydrates, 17% protein, 1.25% cholesterol), and a high-fat, high-cholesterol diet supplemented with ezetimibe (30mg/kg) (HFHC+Z). Mice were fed ad libitum. After two weeks, we injected 106 LNCaP cells into the flanks, with 5-10 mice/group serving as non-injected controls. At two weeks post-injection, half the injected and control mice in the HFHC diet group had ezetimibe added to their diet (HFHC+delayed Z). Tumor volumes and body weights were measured twice weekly. Mice were sacrificed once tumors reached 1000mm3. Body weights and tumor volumes were compared across groups using Kruskal-Wallis test. Interim survival outcomes for this on-going study up to 86 days post-injection were evaluated with Cox proportional hazards.

Results: At 1 day prior to injection, there were no significant differences in body weights across groups. At post-injection day (PD) 2, HFHC+Z mice with tumors were significantly lighter vs. all other mice (p=0.009). By PD 21, HFHC+delayed Z mice with tumors also became significantly lighter vs. the HFLC and HFHC mice with tumors (p<0.001). At PD 86, all mice with tumors were significantly lighter than their control counterparts without tumors (all p<0.013). Tumor volumes were not significantly different across groups (p=0.280). Likewise, survival was not significantly different across groups at PD 86 (p=0.125).

Conclusion: Preliminary evidence from this on-going study demonstrates that ezetimibe is associated with weight loss, but only in mice with PC xenografts. The administration of ezetimibe as preventive vs. treatment therapy does not appear to affect tumor growth or survival. Future studies to explore the interplay between cholesterol metabolism and PC include comparing serum levels of cholesterol, insulin, IGF-1, and IGFBP-3, and tumor markers of proliferation and angiogenesis including akt, pakt, mTOR, VEGF, HIF, CD31 (microvessel density) and ki67 (proliferation). Moreover, we are continuing to follow the mice for any differences in tumor growth or survival.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4602. doi:10.1158/1538-7445.AM2011-4602
Abstract 4619: Chemopreventive activities of the dark-color fruit ‘jamun’, the Indian blackberry
Authors: Gupta Akash , Aqil Farrukh , Kausar Hina , Munagala Radha , Jeyabalan Jeyaprakash , Sharma Ramjee , Singh Inder P. , Gupta Ramesh C.
Published: June 2012

Colored fruits, particularly berries, are emerging as highly chemoprotective because of their antioxidant, anti-proliferative and anti-inflammatory activities. In this study, we investigated the chemoprotective potential of Eugenia jambolana Lam., commonly known as ‘jamun’ or Indian blackberry. Jamun, a widely available fruit in India and other Asian countries, has also been used in the treatment of various ailments, including diabetes mellitus. To determine its antioxidant potential, jamun pulp was collected from freshly harvested fruits, dried, and the powder was extracted with acidic ethanol, and enriched by amberlite XAD7/HP20 (1:1) column chromatography. The pulp powder was found to contain 0.54% anthocyanins, 0.17% ellagic acid/ellagitannins and 1.15% total phenolics using standard assays. HPLC analysis of acid-hydrolyzed extract showed the presence of five anthocyanidins – malvidin, petunidin, delphinidin, cyanidin and peonidin. The unhydrolyzed extract was found to have significant antioxidant activity as reflected by high ORAC value (1,445±64 μmol of TE/g) as well as high ABTS- and DPPH-scavenging activities, and ferrous ions-chelating activity; the acid-hydrolyzed extract showed higher activity than unhydrolyzed extract. Since some of the jamun bioactives (ellagitannins and anthocyanidins/anthocyanins) have significant antiproliferative activity, we also analyzed these extracts for their antiproliferative potential using MTT assay against human lung cancer A549 cells in cell culture. The hydrolyzed jamun extract showed significant antiproliferative activity (IC50=79 µg/mL). However, unhydrolyzed extract showed almost no activity. These data suggest that the antiproliferative activity may result from anthocyanidins (present in the hydrolyzed extract) rather than anthocyanins (present in unhydrolyzed extract). Together, our data show high antioxidant, and antiproliferative activities in jamun pulp and may be a viable candidate for chemoprevention of lung cancer (Supported from Agnes Brown Duggan Endowment).

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4619. doi:10.1158/1538-7445.AM2011-4619
Abstract 4620: Development of honokiol for the prevention of UVB-induced skin tumor development
Authors: Chilampalli Shivani , Averineni Ranjith , Xie Jiashu , Perumal Omathanu , Dwivedi Chandradhar
Published: June 2012

Skin cancer is the most prevalent of all cancers and its incidence is expected to increase substantially. UVB is major etiologic factor in the skin cancer development. Honokiol, a plant ligan isolated from bark and seed cones of Magnolia officinalis, has been shown to have chemopreventive effects on UVB-induced skin cancer development in our laboratory. The objectives of this investigation are to study chemopreventive effects of dose response of honokiol on UVB-induced skin tumor development in SKH-1mice and to prepare topical formulations of honokiol that deliver effective concentration of drug locally. Female SKH-1 mice were divided into 4 groups. Group 1 received acetone one hour before UVB treatment; Group 2, 3 and 4 received honokiol (0.2 ml of 30, 45 and 60 μg in acetone, topical) before each UVB treatment. Mice were treated with UVB dose of 30 mJ/cm2/day for 25 weeks. Tumor counts and group weights were taken weekly. Honokiol pretreated groups resulted in 36-78% reduction in tumor multiplicity compared to control. Honokiol pretreatment (60 μg) decreased tumor incidence by 40 %. Topical formulation of honokiol gel was prepared by using carbomer 940 as base and propylene glycol as penetration enhancer. Drug content and content uniformity were determined. Release studies and permeation studies were studied in detail.

This study is supported by Translational Cancer Research Center funded as 2010 Research Initiative Center by the State of South Dakota

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4620. doi:10.1158/1538-7445.AM2011-4620
Abstract 4622: Determination of pyranocoumarin compounds decursin, decursinol angelate and decursinol simultaneously in mouse plasma and tumor tissue by liquid-liquid extraction and HPLC
Authors: Li Li , Zhang Jinhui , Zhang Yong , Shaik Ahmad Ali , Wang Lei , Xing Chengguo , Kim Sung-Hoon , Lü Junxuan
Published: June 2012

Pyranocoumarin compound decursin and its isomer decursinol angelate (DA) are the major chemical components of the root of Angelica gigas Nakai (AGN, Korean Angelica), a major traditional medicinal herb. Recently, AGN has been shown to exhibit anti-tumor activities by our collaborative team. Decursinol has been identified as a major metabolite of decursin and DA in vivo by us and others, but another group has published very controversial results. To better define the mechanisms of in vivo action for AGN, pharmacokinetic studies are essential. In this study, we set up a practical method for the concurrent analysis of decursin, DA and decursinol in mouse plasma and tumor tissues by liquid-liquid extraction and HPLC-UV. In brief, plasma or tumor homogenate (0.1-0.2 ml) spiked with 1μg chrysin as internal standard (IS) was extracted directly with ethyl acetate. After extraction, decursin, DA and decursinol were quantitated by HPLC with UV detection at 330 nm. The method was validated in terms of recovery, lower limit of quantitation and linear range. The extraction efficiency for spiked decursin/DA and decursinol at 1 µg/ml was 90.1±3.2 % and 90.3±1.8% in mouse plasma, respectively. The lower limit of quantitation (LOQ) was approximately 0.25 μg/mL for decursin/DA, and 0.2 μg/mL for decursinol in mouse plasma. The linear range of the method was 0.25 -25 µg/ml and 0.2-50 µg/ml in mouse plasma for decursin/DA and decursinol, respectively.

In a pilot pharmacokinetic study, male C57BL/6 mice were given a single dose of 4.8 mg decursin/DA (∼240mg/kg) per mouse either by gavage or intraperitoneal injection in 0.2 mL vehicle of ethanol:PEG400:Tween80:5% glucose (3:6:1:20). Maximum plasma concentrations for decursin/DA and decursinol were found to be 34.2 and 324 µM respectively when D/DA was administered via intraperitoneal injection and to be 1.7 and 60.7 µM via gavage. These findings further confirmed that decursinol was one of the major metabolites of decursin/DA in vivo. In athymic nude mouse xenograft study, the mice were given intraperitoneal injection of decursin/DA at a dose of 1mg (∼40mg/kg) daily, 5 days per week, for 7 weeks. The decursin/DA and decursinol concentration was 1.5 and 10μg/g, respectively, in the tumor tissue.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4622. doi:10.1158/1538-7445.AM2011-4622
Abstract 4626: Chamomile confers protection against hydrogen peroxide-induced toxicity through activation of Nrf2-mediated defense response
Authors: Bhaskaran Natarajan , Shukla Sanjeev , Gupta Sanjay
Published: June 2012

Cellular damage by reactive oxygen species (ROS) causes aging and remains major factor in the development of various human diseases including diabetes, liver diseases, neurodegenerative and cardiovascular diseases, including cancer. Dietary antioxidants have been considered as an attractive strategy to prevent or attenuate the progression of diseases caused by oxidative stress. Induction of phase 2 detoxifying enzymes in response to naturally occurring agents provides an effective means of protection against oxidative stress. Aqueous extract of chamomile (Matricaria chamomilla) in the form of tea has been used for centuries to treat various inflammatory conditions supporting its traditional use for treating various human ailments. We used aqueous chamomile extract obtained from dried flowers to investigate its effect on Nrf2 activation and induction of phase 2 enzymes in RAW 264.7 murine macrophages and H2O2 as an agent for oxidative stress. Treatment with 50 µM H2O2 for 6 h caused significant increase in cellular stress accompanied by cell death in RAW 264.7 murine macrophages as evident from light microscopy and annexin-V FITC assay. Pre-treatment with chamomile at 10, 20 and 40 µg for 16 h followed by H2O2 treatment protected cell death caused by excessive ROS generation and prevented H2O2-mediated inhibition of cell growth. Chamomile exposure significantly increased the expression of antioxidant enzymes viz. peroxiredoxin (Prx), thioredoxin (Trx) and heme oxygenase (HO-1) in a dose-dependent manner, compared with their respective controls. Furthermore, chamomile exposure to RAW 264.7 murine macrophages increased the nuclear translocation of Nrf2 with increased phosphorylated Nrf2 levels, and it's binding to the antioxidant response element (ARE) in the nucleus. These data provide evidence that chamomile induces the expression of cytoprotective antioxidant enzyme (HO-1) and other redox regulatory enzymes (Prx and Trx) in response to the nuclear translocation of a key redox sensitive transcription factor Nrf2 from the cytoplasm and provide protection from cytotoxicity caused by inflammatory oxidants. Understanding chamomile-mediated molecular events could play a potentially important role in setting up strategies for prevention and treatment of various human diseases including cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4626. doi:10.1158/1538-7445.AM2011-4626
Abstract 4627: Potent chemotherapeutic activity of green tea polyphenols, blueberry anthocyanidin mixture and cisplatin: Synergistic effects
Authors: Cao Pengxiao , Kausar Hina , Gupta Ramesh
Published: June 2012

A significant progress has been made in the prognosis of several cancers (e.g., breast and prostate cancers). However, the prognosis of lung cancer remains grim. Furthermore, up to 50% of stage I/II non-small-cell lung cancer (NSCLC) patients develop recurrence/metastasis within five years of their treatments. It is eminent that natural compounds or food-based bioactives with minimal toxicity be identified for their use in neo-adjuvant chemotherapy and/or after the primary cancer treatment for enhanced efficacy. Green tea and berries are drawing increased attention due to their potential anti-cancer effects. The active compounds in green tea and berries are polyphenols, e.g., epigallocatechin gallate (EGCG) and anthocyanidins (e.g. delphinidin, cyanidin, peonidin, petunidin, malvidin in blueberry), respectively. We previously demonstrated synergistic antiproliferative activity from these Anths against H1299 lung cancer cells both in vitro and in vivo (Kausar et al. AACR 51: 1884, 2010). In this study we investigated the combined effect of EGCG and the blueberry anthocyanidins (EGCG-Anths) alone and in combination with a common chemotherapeutic drug, cisplatin on lung cancer H1299 cell using MTT assay. The antiproliferative activity of EGCG-Anths (6 μM), measured as IC50, was found to be 2- and 4-fold lower compared with Anths (12 μM) and EGCG (21 μM) alone, respectively. The IC50 of cisplatin was found to be ≈50 μM. However, when combined with the EGCG-Anths (10 μM each), the IC50 of cisplatin was significantly reduced (<3 μM). Flow cytometric analysis for cell cycle distribution and apoptosis revealed cell cycle arrest and enhanced apotosis induced by the combination mixture. Western blot analysis for the involved molecular targets showed that selected proteins (cyclin D1, B1, Cdc2, p21, PCNA, Bax, Bcl-2, PARP, etc.) associated with cell cycle and apoptosis were more significantly modulated by the combination, EGCG-Anths-cisplatin than with the individual entities. Together, our data suggest that combination of EGCG-Anths alone or in combination with suboptimal doses of cisplatin may be effective in neo-adjuvant chemotherapy or can be used as maintenance drug for the prevention of lung cancer recurrence/metastasis and minimize toxicity issues associated with high dose cisplatin (Supported from KLCRP, T32-ES011564 and Agnes Brown Duggan Endowment).

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4627. doi:10.1158/1538-7445.AM2011-4627
Abstract 4630: Cruciferous vegetable glucosinolate hydrolysis products protect against Benzo[a]pyrene-induced genotoxicity
Authors: David Rhiannon M. , Patel Saroor , Abd Kadir Nurul H. , Rossiter John , Gooderham Nigel
Published: June 2012

Recent epidemiology studies have indicated that cruciferous vegetable intake is inversely correlated with lung cancer risk among smokers (Tang et al., BMC Cancer 2010, 10, 162). Benzo[a]pyrene (BaP) is a prominent contaminant of cigarette smoke, a model genotoxic carcinogen and powerful mutagen. Glucosinolates found in cruciferous vegetables are known to be hydrolysed to compounds that have been shown to be chemopreventive in both animal and cell-based systems, while, paradoxically, evidence is gathering that high concentrations of some of these hydrolysis products are genotoxic.

The aim of the current study was to investigate whether pre-treatment of MCL-5 cells (a metabolically competent human lymphoblastoid cell line) with glucosinolate hydrolysis products conferred protection against the cytotoxicity and genotoxicity induced by a subsequent exposure to BaP. In the case of AITC (allyl isothiocyanate), a model glucosinolate hydrolysis product, toxicity and mutagenicity to MCL-5 cells was established for a range of concentrations. Mutagenicity was assessed through resistance to trifluorothymidine at the thymidine kinase (TK) locus and 6-thioguanine at the hypoxanthine-guanine phosphoribosyltransferase (HPRT) locus. The chemical treatments revealed a consistent trend for reduced cytotoxicity in cells pre-treated with AITC prior to BaP compared to those treated with BaP alone. Mutation frequency at the TK locus was consistently lower in the cells pre-treated with AITC compared to those treated with BaP alone. High concentrations of AITC induced a prolonged, dose-dependent cytotoxicity with cells only returning to normal growth rate after 7 days.

These data suggest that pre-treatment of MCL-5 cells with glucosinolate hydrolysis products such as AITC affords protection against the genotoxic insult induced by mutagens such as BaP.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4630. doi:10.1158/1538-7445.AM2011-4630
Abstract 4637: Intra-individual changes in circulating IGF-1 and IGF-1/IGFBP-3 molar ratio are associated with colorectal adenomatous polyps presence
Authors: Soubry Adelheid , Sedjo Rebecca , Wang Frances , Byers Tim , Rosen Clifford , Yashin Anatoli , Ukraintseva Svetlana , Haffner Steve , D'Agostino Ralph , Il'yasova Dora
Published: June 2012

Background: High levels of circulating insulin-like growth factor-1 (IGF-1) have been associated with increased risks of several cancers, including colorectal cancer.

Methods: In a sub-cohort of the Insulin Resistance and Atherosclerosis Study (IRAS), we examined the associations between circulating IGF-1 and IGF-1/IGFBP-3 molar ratios, as well as their change patterns during 10-year follow-up, with the presence of colorectal adenomatous polyps at the end of the follow-up.

Results: The general trend was decrease of both variables with age. Based on this trend, two patterns were considered: “no increase” (variation within 15% of baseline and/or decrease) and “ever increase” (at least one increase of > 15%). In the subgroup of participants with normal glucose tolerance, “ever increase” was positively associated with colorectal adenoma: ORs were 3.65 (1.30-10.8) and 2.83 (1.00-8.22) for IGF-1 and IGF-1/IGFBP-3, respectively. No association was found between actual levels of IGF-1 or IGF-1/IGFBP-3 at any time point and the presence of colorectal adenoma.

Conclusion: Our data suggest that increase in circulating IGF-1 and IGF-1/IGFBP-3 may represent an unnatural trend of a disturbed signaling pathway, which could favor the development of precancerous lesions, such as colorectal adenoma.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4637. doi:10.1158/1538-7445.AM2011-4637
Abstract 4642: Biomarkers for assessment of tissue quality in cancer research
Authors: Li Jie , Eason Arianne , Vortmeyer Alexander
Published: June 2012

Objectives: Much of the progress in molecular cancer research would not have been achieved without access to clinical tissue specimens. However, lack of quality control on these specimens frequently leads to the generation of irreproducible research data, especially when archived samples of uncertain quality are utilized. We hypothesized that biomolecules have different susceptibility to decomposition and that susceptible molecules may serve as indicators to reveal the molecular quality of tissue. We seek to use a proteomic approach to evaluate protein decomposition over time at room temperature (RT) and to mathematically coordinate the extent of decomposition with the RT-exposure time length. This decomposition curve will serve for tissue-quality assessment.

Materials and methods: Cultured cells were used for primary biomarker-screening. Cell origins included kidney (293T), prostate (LNCaP), breast (MDA-MB-231), blood (Jurkat), and lymphoma (Namalwa). Aliquots of cells were randomly assigned to immediate flash-freezing or exposure to RT for 0.5, 4, 8, 24, 48 hours. Lysates were applied in duplicate to Two-Dimensional Fluorescence Difference Gel Electrophoresis (2D DIGE) with pH 3-10 IPG strips and large format PAGE. Proteomic comparison was performed using Decyder software with embedded paired t-/ANOVA-analysis. Spots, consistent in repeats with statistical significance between groups, were sequenced with both LC/MS/MS and MALDI-TOF. Clinical specimens (n=3 for each corresponding organs), procured as above, were used for biomarker validation via regular Western blot and 2-D/IPG-strip Western blot.

Results: 2D-DIGE revealed ∼3000 spots per gel. During the course of exposure to RT, proteomic patterns exhibited dynamic and time-dependent changes in specific subsets of proteins, starting from 0.5 hour. The alterations revealed on 2D-DIGE were changes in molecular weight and/or isoelectric point; the results were validated via Western blot and 2-D/IPG-strip Western blot, respectively. Protein candidates that decomposed in a time-dependent manner allowed for the generation of regression curves using the protein quantity against corresponding time points. Several of above proteins, mostly housekeeping proteins, behaved consistently across all types of observed cell/specimen. Therefore, they could serve as widely applicable biomarkers for indication of exposure time (to RT) and further for assessment of the molecular quality of research specimens.

Conclusions: The study will draw peers’ attention on the molecular quality of their research tissue and offer biomarkers for the quality assessment. Based on the assessment result, the meaning of tissue-research data should be reinterpreted accordingly.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4642. doi:10.1158/1538-7445.AM2011-4642
Abstract 4652: Urinary levels of the tobacco-specific carcinogen N′-nitrosonornicotine and its glucuronide are strongly associated with esophageal cancer risk in smokers in the Shanghai Cohort Study
Authors: Yuan Jian-Min , Knezevich Aleksandar D. , Wang Renwei , Gao Yu-Tang , Hecht Stephen S. , Stepanov Irina
Published: June 2012

Background: Experimental studies have shown that the tobacco-specific nitrosamine N′-nitrosonornicotine (NNN) can induce esophageal cancer in rats. However, epidemiological data that directly link NNN to esophageal cancer in humans have been lacking. Methods: A nested case-control study of esophageal cancer including 77 cases and 223 individually matched control subjects within the Shanghai Cohort Study was conducted to examine the association between urinary NNN and risk of developing esophageal cancer among male current smokers. Besides urinary creatinine (Cr) and total cotinine, urinary free NNN and its pyridine-N-glucuronide (NNN-N-Gluc) were quantified separately for each study sample by liquid chromatography- tandem mass spectrometry. A conditional logistic regression method was used to calculate the odds ratios (ORs) and 95% confidence intervals (CIs) for esophageal cancer associated with elevated urinary NNN. All P values reported were two-sided. Results: Esophageal cancer patients had significantly higher urinary levels of free NNN (geometric mean 58.2 fmol/mg Cr), NNN-N-Gluc (49.8 fmol/mg Cr) and the sum of the two, referred to as total NNN (117.9 fmol/mg Cr), than control subjects (geometric means 18.2, 24.4, and 45.1 fmol/mg Cr, respectively) whereas the ratio of NNN-N-Gluc to total NNN was significantly lower in cancer patients (46.7%) than controls (56.4%) (all P's < 0.0001). Compared with the lowest quartile, ORs (95% CIs) for esophageal cancer for the 2nd, 3rd, and 4th quartiles of free NNN were 2.61 (0.80-8.55), 5.74 (1.67-19.75), and 30.64 (7.70-121.93), respectively, after adjustment for smoking intensity and duration (Ptrend < 0.0001). Additional adjustment for urinary cotinine strengthened the association, and ORs (95% CIs) became 4.46 (1.14-17.41), 13.83 (3.21-59.53) and 128.22 (19.9-826.7) (Ptrend < 0.0001). In contrast, the ratio of NNN-N-Gluc to total NNN was inversely associated with esophageal cancer risk. The smoking-adjusted ORs (95% CIs) for the 2nd, 3rd, and 4th quartiles were 0.43 (0.20-0.96), 0.23 (0.09-0.56) and 0.20 (0.08-0.50), respectively, compared with the lowest quartile (Ptrend =0.0002). Additional adjustment for urinary total cotinine did not materially alter the observed ORs. Conclusion: The findings of the present study indicate that the tobacco-specific nitrosamine NNN plays a significant role as a cause of esophageal cancer in smokers. In contrast, the glucuronidation of NNN may protect against the development of tobacco-related esophageal cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4652. doi:10.1158/1538-7445.AM2011-4652
Abstract 4662: Genetic predictors of clinical outcomes for patients with non-small cell lung cancer
Authors: Pu Xia , Spitz Margaret R. , Amos Chris , Ye Yuanqing , Hildebrandt Michelle A. , Chang David , Lu Charles , Stewart David , Roth Jack A. , Lippman Scott M. , Christiani David , Wu Xifeng
Published: June 2012

Background: Emerging evidence has shown that genetic factors can interact with host factors to influence patient's clinical outcomes. Inflammatory biomarkers have been identified as prognostic factors independent of tumor stage. To test the hypothesis that genetic variants in inflammation-related genes may modulate clinical outcome in non-small cell lung cancer (NSCLC) patients, we conducted a three-stage study, including an initial discovery phase and two validation phases. Methods: In the discovery phase, we analyzed the association between 19,458 SNPs in inflammation-related genes and SNPs from GWAS risk analysis, and clinical outcomes in 1,408 NSCLC patients. In the validation phases, the significant SNPs (p<0.05) were first validated in an additional 1,104 NSCLC patients from MD Anderson and then validated in 978 NSCLC patients from Harvard University. Meta-analysis was used to summarize the combined effect of each SNP in all three stages. Results: A total of 16 SNPs were identified as top candidate predictors of respective outcomes after showing consistent associations in all three stages. Among them, 5 SNPs were associated with overall survival in the entire populations. The most significant SNP is located on chromosome 6 (discovery: HR=1.49, 95%CI=1.10-2.01, P=0.010; combined validation: HR=1.45, 95%CI=1.17-1.78, P=0.001; all combined: HR=1.46, 95% CI=1.23-1.73, P=1.59×10-5). Five SNPs were associated with overall survival in late stage unresectable patients receiving chemotherapy. The most significant SNP, located on chromosome 13, consistently showed increased risk of death in all three phases (discovery: HR=1.40, 95%CI=1.09-1.81, P=0.010; combined validation: HR=1.36, 95%CI=1.14-1.63, P=0.001; all combined: HR=1.37, 95% CI=1.19-1.59, P=2.26×10-5). Six SNPs were consistently associated with progression in early stage patient receiving surgery only. The top SNP was on chromosome 1 with protective effect against developing progression in all three phases (discovery: HR=0.52, 95%CI=0.32-0.86, P=0.033; combined validation: HR=0.63, 95%CI=0.49-0.82, P=0.001; all combined: HR=0.62, 95% CI=0.5-0.79, P=5.47×10-5). To our knowledge, this is the first study to systematically evaluate the association of genetic variations in major inflammation genes and NSCLC patients’ clinical outcomes with a multi-stage design using two of the largest lung studies in the US. Our study provides evidence that genetic variations in inflammation genes may affect clinical outcomes in NSCLC patients.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4662. doi:10.1158/1538-7445.AM2011-4662
Abstract 4663: Genetic variation in the p38 signaling pathway as markers for postoperative pulmonary complications in lung cancer patients
Authors: Hildebrandt Michelle A. , Roth Jack A. , Vaporciyan Ara A. , Pu Xia , Ye Yuanqing , Correa Arlene M. , Kim Jae Y. , Swisher Stephen G. , Wu Xifeng
Published: June 2012

Surgical resection is the primary treatment for patients with localized non-small cell lung cancer (NSCLC). Although often a successful treatment option, a portion of these individuals will experience postoperative pulmonary complications, such as pneumonia, acute respiratory distress syndrome, and atelectasis. These complications are difficult to predict and often characterized by a hyper-inflammatory response. The p38 signaling pathway is activated following stress stimuli and in response to inflammatory signals. Because of this important role, we hypothesized that genetic polymorphisms within the core components of this pathway may result in dysregulated response to inflammation following surgery causing unforeseen complications. Therefore, in this study we genotyped 242 common, germline genetic variants from 19 p38 pathway genes in 275 Caucasian NSCLC patients who underwent a lobectomy at MD Anderson. After controlling for clinical risk factors, twenty-three significant associations with any pulmonary complication were observed (P<0.05). Patients carrying at least one variant allele of IRAK2:rs778044 had a complication rate of 49.7% compared to 28.6% for patients with the common genotype for a highly significant 3.10-fold increase in risk (95% Confidence Interval [CI]: 1.76-5.47, P=9.6×10-5). A dose-response effect was evident between number of risk genotypes and risk of complication (P=2.08×10-7). Patients with the highest number of risk genotypes were nearly 14 times more likely to develop a pulmonary complication (95% CI: 5.30-36.87, P=9.73×10-8) compared to those with the fewest risk genotypes. Potential gene-gene interactions were also observed between IRAK2, MYD88, and IL1R1 that identified distinct risk groupings. Patients carrying both the variant genotypes for IRAK2:rs778044 and MYD88:rs11783456 were at highest risk, while those with the common genotype for IRAK2:rs778044 and the protective variant genotype for IL1R1:rs2287047 had the lowest risk. The increase in risk was 23.57-fold for high risk individuals compared to those with the low risk genetic profile (95% CI: 2.75-201.96, P=0.0039). These results strongly implicate the p38 pathway in modulating risk of pulmonary complications following lobectomy for NSCLC. Together with clinical and epidemiological variables, these variants may be useful biomarkers to better identify patients at high risk prior to surgery.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4663. doi:10.1158/1538-7445.AM2011-4663
Abstract 4670: Polymorphisms in DNA repair genes, chemotherapy and survival in bladder cancer
Authors: Ricceri Fulvio , Sacerdote Carlotta , Polidoro Silvia , Guarrera Simonetta , Allione Alessandra , Critelli Rossana , Critelli Rossana , Russo Alessia , Gontero Paolo , Destefanis Paolo , Matullo Giuseppe , Vineis Paolo
Published: June 2012

Bladder cancer (BC) survival is highly influenced by environmental and predisposing genetic factors. In the last decades growing evidence of the major role played by DNA repair systems in the survival after a BC has been provided.

The objectives of the study are to assess the relationship between DNA Repair Genes polymorphisms and survival by treatment in bladder cancer patients.

We conducted a hospital-based case-control investigation at S. Giovanni Battista hospital in Turin, where about half of the incident BC in the Turin metropolitan area are treated. Cases are histologically confirmed incident BC patients, male, aged 40 to 75 years and resident in the Turin metropolitan area. Each patient filled in a questionnaire on history of tobacco smoking, occupational history, 24-hours medication use recall and food habits, and provided a blood sample. We followed up BC patients for death and recurrences using municipal registry office, death's certificate, hospital discharge, medical records and pathologists’ reports. In the cohort of BC patients, we genotyped 36 Single Nucleotide Polymorphisms (SNPs) in 10 DNA Repair Genes.

We analyzed survival on BC patients using a Cox proportional-hazards regression model for codominant and per-allele scenario, adjusted for age, stage and grading and stratified by therapy (chemotherapy vs no or other therapies). Chemotherapy group include patients tretated with systemic and local chemotherapy with different therapeutic plans (Carboplatinum doublets, Gencitabin, Epirubicin, etc).

We identified 456 BC patients which were followed-up for mortality (median follow-up 62 months). In multivariate analysis both stage and grade predicted the outcome in a statistically significant way, after adjustment for age. Our preliminary analyses showed that the variant alleles in two SNPs in XRCC1 gene (rs 2854509 and rs 3213255) confer a significant lower risk of death and one SNP in ERCC2 gene (rs 171140) confers a significant higher risk of death compared to the wild type (analyses adjusted by age, stage and grading) only in the group of treated with chemotherapy.

Our findings suggest a link between DNA repair genes variation and survival in BC patients that become evident only in patients treated with chemotherapy. We hypothesize that levels of DNA adducts or other DNA damages inducted by chemotherapy, are partially due to the individual DNA repair activity, resulting in a different response to chemotherapy and, consequentially, in different survival rates.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4670. doi:10.1158/1538-7445.AM2011-4670
Abstract 468: Down-regulation of TNFSF15 (VEGI) in tumor vasculature under inflammatory conditions is a pre-requisite of tumor neovascularization
Authors: Deng Weimin , Lu Yi , Gu Xin , Liang Paulina Huang , Yao Zhi , Li Luyuan
Published: June 2012

Tumor necrosis factor superfamily-15 (TNFSF15; VEGI) is an endogenous inhibitor of neovascularization. TNFSF15 is largely produced by quiescent vascular endothelial cells in an established vasculature but is markedly down-regulated in endothelial cells undergoing proliferation such as those in tumors or at a wound site. Recombinant TNFSF15 is a potent inhibitor of endothelial cell proliferation and tumor growth. We hypothesize that TNFSF15 as a negative regulator of angiogenesis must be down-regulated prior to the initiation of angiogenesis, and that the inflammatory conditions created by cancer cells and infiltrating lymphocytes lead to TNFSF15 down-modulation in the microenvironment, permitting new blood vessel growth. We analyzed normal ovary tissues (n=12) and clinical specimens of ovarian cancer (n=86; including stages I, II, III and IV) for the expression of TNFSF15, the endothelial cell marker PECAM (CD31), and tumor-infiltrating T-cell markers CD4 and CD25. We found that the degree of T-cell infiltration increases markedly while TNFSF15 expression diminishes as the disease progresses. We then determined the influence of human ovarian cancer cell line OVCAR-3 cell conditioned media on cytokine production by human CD4+ T-cells and CD4+CD25+ Treg cells, as well as the influence of the conditioned media of the latter two cells on TNFSF15 expression in human umbilical vein endothelial cells (HUVEC). We found that the conditioned media of Th1 or Treg cells can inhibit HUVEC TNFSF15 expression only after the lymphocytes being treated with OVCAR-3 conditioned media. Interestingly, we further found that mouse ovarian cancer ID8 cells implanted subcutaneously on mice treated with TNFSF15 shRNA at the injection sites exhibit significantly greater tumor growth rates compared with control shRNA treated ones. Our findings indicate that TNFSF15 plays a critical role in the maintenance of vascular stability as its down-modulation is a pre-requisite for the initiation of tumor neovascularization, and that TNFSF15 down-regulation is carried out by tumor-infiltrating T-cells under the influence of the cancer cells in the microenvironment.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 468. doi:10.1158/1538-7445.AM2011-468