bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3699: Novel combinatorial strategies for cancer chemoprevention using polymethoxyflavones from orange and atorvastatin
Authors: Qiu Peiju , Charoensinphon Noppawat , Zheng Jinkai , Song Mingyue , Dong Ping , Riddle Ryan , Xiao Hang
Published: June 2012

Cancer is a complex disease, and multiple aberrant molecular events are driving forces for cancer progression. It is difficult to control cancer progression by a single agent. We investigated a novel approach to inhibit cancer cell growth by using polymethoxyflavones (PMFs, isolated from aged orange peel) and atorvastatin (Lipitor, a commonly used cholesterol-lowering drug) in combination. The goal is to identify combination regimen that can produce synergy in inhibiting human cancer cell growth. Our results on multiple colon and lung cancer cells demonstrated that permethoxylated PMFs, namely nobiletin and tangeretin, can synergistically interact with atorvastatin and produced much stronger inhibitory effects than either agent alone. On the other hand, the hydroxylated PMFs, namely 5-hydroxy nobiletin (5HN) and 5-hydroxy tangeretin (5HT) showed different interaction profiles with atorvastatin, i.e., 5HN produced strong synergy with atorvastatin while 5HT did not. Further mechanistic study demonstrated that the combination treatment with 5HN and atorvastatin for 24 hr caused extensive cell population accumulation in G0/G1 phase; whereas at 48 hr or longer, apoptosis was induced significantly. Our results also demonstrated that the combinational effects of 5HN and atorvastatin were associated with increased levels of p21Cip1/Waf1, and p27Kip1; decreased levels of CDK-2, CDK-4,CDK-6,Cyclin D1, Mcl-1 and hyper-phosphorylated Rb(Ser 780); and activation of caspase cascade. More importantly, we investigated the inhibitory effects of PMFs/atorvastatin combinations in an NNK-induced mouse lung tumorigenesis model, and our results demonstrated that dietary administration of nobiletin and 5HN in combination with atorvastatin caused potent inhibition of lung tumor formation in A/J mice. These inhibitory effects were much stronger than those produced by singular nobiletin, 5HN, or atorvastatin treatments at much higher doses. The isobologram-based statistic analysis confirmed that nobiletin, 5HN and atorvastatin produced strong synergy in inhibiting lung tumorigenesis in mice. In conclusion, the present work provided promising leads for future investigation on the novel combination of PMFs and atorvastatin as novel strategy for cancer chemoprevention. (This study was supported in part by NIH-CA139174, AICR-#10A044, CEAR pilot grant, and USDA Special Grant on Bioactive Foods).

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 3699. doi:10.1158/1538-7445.AM2011-3699
Abstract 3701: Dimethylbenzanthracene (DMBA) and DMBA dihydrodiol mutagenicity in rat epithelial and fibroblast cell lines, and its inhibition by combinations of nutraceuticals
Authors: Sacks Peter G. , Zhao Zhong-Lin , Kosinska Wieslawa , He Zhiming , Josephy P. David , Guttenplan Joseph B.
Published: June 2012

7,12-Dimethylbenzanthracene (DMBA) is a potent mammary carcinogen in rats. Combinations of non-toxic nutraceutical agents, administered at or near physiological levels, were investigated for their abilities to inhibit the mutagenicity of DMBA or DMBA-dihydrodiol (DMBAD, a primary metabolite and proximate mutagen of DMBA) in vitro, in rat mammary epithelial and fibroblast cells derived from a lacI (BigBlue) Fischer rat (McDiarmid, H.M., Douglas, G.R., Coomber, B.L., and Josephy, P.D. Epithelial and fibroblast cell lines cultured from the transgenic BigBlue rat: an in vitro mutagenesis assay. Mutat. Res., 497: 39-47, 2001). In the epithelial cells, DMBA was not appreciably mutagenic at concentrations up to 4 µM, but DMBAD, was significantly mutagenic at ten-fold lower concentrations. These results indicate that the epithelial cells can bioactivate the intermediate, DMBAD, but cannot effect the complete biotransformation of DMBA to its ultimate mutagenic metabolite, DMBA-dihydrodiolepoxide. In the fibroblast cell line, in contrast, DMBA was mutagenic at concentrations as low as 20 nM and DMBAD was even more potent than DMBA. Several combinations of nutraceuticals (dietary components providing health benefits) were tested for their abilities to inhibit mutagenesis in these cell lines; the concentrations tested were based on reported serum concentrations and these were used to establish 1x concentrations. The agents and their 1x concentrations were: resveratrol (Res), 2.4 μM; sulforaphane (Sul), 0.06 μM; antioxidant mix (α- and γ-tocopherol, 30 μM, plus vitamin C, 68 μM – VCE); α-lipoic acid (LA), 2 μM; epigallocatechin gallate (EGCG), 0.7 μM; and N-acetylcysteine (NAC), 12 μM. None of the agents or their combinations, tested at 1 – 3 x concentrations, showed any cytotoxicity. In both epithelial and fibroblast cells, Sul and Res alone slightly inhibited mutagenesis at 2x concentrations; no other agents had observable effects. All binary combinations of Res, LA, and Sul, at 2x concentrations, inhibited mutagenesis; the Res + Sul combination was particularly effective (ca. 50% inhibition). These results suggest a role for fibroblast cells in the bioactivation of carcinogens, implicating the microenvironment, and indicate that combinations of nutraceuticals can inhibit mutagenesis by polycyclic aromatic hydrocarbons. Supported by the Susan Komen Foundation grant # KG080836 (JBG) and NSERC Canada (PDJ).

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 3701. doi:10.1158/1538-7445.AM2011-3701
Abstract 3704: Diindolylmethane induces ER stress mediated autophagy in ovarian cancer cells
Authors: Kandala Prabodh K. , Srivastava Sanjay K.
Published: June 2012

Endoplasmic reticulum (ER) is responsible for biosynthesis, folding assembly and modification of various soluble and membrane proteins. ER stress is the imbalance in protein folding resulting in the accumulation of misfolded proteins in ER lumen. In our study, we observed that Diindolylmethane (DIM), a compound present in cruciferous vegetables induces ER stress in ovarian cancer cells. Treatment of SKOV-3, OVCAR-3 and TOV-21G ovarian cancer cells with varying concentrations of DIM for 24hours resulted in the upregulation of Bip, a molecular chaperone. Bip or Grp78 binds to IRE1α, ATF6 and PERK under normal condition, however, during ER stress, Bip dissociates and releases all the three proteins. DIM treatment also significantly increased the expression of IRE1α, ER oxidase (ERO1) and CHOP. CHOP translocate to nucleus and initiate apoptotic stimuli in cells undergoing ER stress. Moreover, DIM induced ER stress also lead to the activation of eIF2α by phosphorylation. Activation of eIF2α inhibits protein translation to avoid further protein accumulation in ER. The release of calcium by DIM treatment in ovarian cancer cells was evaluated by Accuri C6 flowcytometer. The advantage with Accuri C6 is that calcium levels can be monitored without interruption. DIM treatment increased the cytosolic calcium levels in SKOV3 cells. Increase in cytosolic calcium is known to cause ER stress. In addition, our flowcytometry results indicate that DIM induces autophagy in all the ovarian cancer cells in a concentration dependent manner. Exposure of SKOV-3, OVCAR-3 and TOV-21G cells to DIM upregulated the expression of LC3B, a potent indicator of autophagy. DIM induced autophagy was blocked by bafilomycin, an autophagy inhibitor. Treatment of cells with mithramycin (ER stress inhibitor) blocked ER stress proteins activated by DIM. Interestingly, we also observed that blocking DIM-induced ER stress by mithramycin blocked autophagy in ovarian cancer cells. Taken together these results suggest that induction of autophagy in ovarian cancer cells was associated with ER stress caused by DIM. Further mechanistic studies are in progress. [Supported in part by R01 grants CA106953 and CA129038 (to S.K.S) awarded by the National Cancer Institute].

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 3704. doi:10.1158/1538-7445.AM2011-3704
Abstract 3717: Evaluation and comparison of breast cancer risk prediction models for Hispanic and non-Hispanic white women in the Women's Health Initiative
Authors: Banegas Matthew P. , Gail Mitchell H. , LaCroix Andrea , Thompson Beti , Martinez M. Elena , Wactawski-Wende Jean , John Esther M. , Hubbell F. Allan , Yasmeen Shagufta , Katki Hormuzd A.
Published: June 2012

Introduction: The National Cancer Institute (NCI) Breast Cancer Risk Assessment Tool (BCRAT) has not been previously validated for use in Hispanic women. We evaluated and compared BCRAT risk projections for Hispanic and non-Hispanic White (NHW) participants in the Women's Health Initiative (WHI).

Methods: We used data on 6,248 Hispanic and 127,426 NHW post-menopausal WHI participants aged 50 years and older. Absolute risks of developing invasive breast cancer over the WHI main and extension studies were calculated using the BCRAT and an updated BCRAT using 1993-2007 SEER breast cancer incidence rates. We assessed model calibration by the ratio of observed to expected number of breast cancers (O/E ratio) and discriminatory accuracy with the concordance statistic (c-statistic).

Results: In the WHI main study, the BCRAT underestimated the number of breast cancers in Hispanics (O/E ratio=1.18, 95% CI = 0.99, 1.40) and NHWs (O/E ratio=1.18, 95% CI = 1.14, 1.21). The updated BCRAT was well calibrated for Hispanics (O/E ratio=1.08, 95% CI = 0.91, 1.28) and NHWs (O/E ratio=0.98, 95% CI = 0.96, 1.01). The c-statistic was 0.629 (95% CI = 0.582, 0.676) for Hispanics and 0.575 (95%CI = 0.566, 0.583] for NHWs. In the WHI extension study, where women reported being screened less frequently, BCRAT overestimated risk in both Hispanics and NHWs.

Conclusion: Underestimation of breast cancer risk by the BCRAT in Hispanic and NHW women was corrected by using updated SEER breast cancer incidence rates. Recalibrating the BCRAT with current SEER incidence rates and racial/ethnic specific relative risk estimates may improve the predictive accuracy for Hispanic women.

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 3717. doi:10.1158/1538-7445.AM2011-3717
Abstract 3723: Predicting breast cancer risk to age 80: A clinical tool
Authors: Colditz Graham A. , Rosner Bernard
Published: June 2012

While numerous models of breast cancer risk have been developed, few are used in routine clinical care. Thus women are not advised of potential strategies to reduce their future risk of breast cancer; either SERMS for high risk women or weight control and physical activity, reduction in alcohol or other approaches for moderate risk women. Furthermore, few models incorporate the full range of known risk factors and provide easy access for clinical use. The Rosner-Colditz log incidence model of breast cancer was developed and validated on independent data sets within Nurses’ Health Study cohort 1. The model includes a full range of reproductive and lifestyle risk factors for breast cancer. We now have developed a tool for batch or individual data input that provides 5-year risk, and risk to age 80. Input variables include current age, weigh at age 18, current weight, height, current alcohol intake and at age 18, age at menarche, age at menopause and type of menopause, family history of breast cancer and history of benign breast disease, hormone use (current, past, never) and type of hormone used, duration of postmenopausal hormone use, and age at each birth. Parameter estimates are drawn from Nurses’ Health Study data where we have shown the model performs with higher predictability (area under the ROC curve was 0.623) significantly better than the Gail model (area under the ROC 0.578) (p<0.001). For the total cohort, Incidence of invasive breast cancer in the NHS is consistent with SEER data (O/E = 1.0; 95%CI 0.98-1.03). For clinical use, risk estimates are adjusted for US mortality to provide incidence estimates to age 80 that account for competing risks. Clinical application requires assumptions regarding future weight change, duration of continued use of hormone therapy, and for premenopausal women, future age at menopause.

1. Colditz G, Rosner B. Cumulative risk of breast cancer to age 70 years according to risk factor status: data from the Nurses’ Health Study. Am J Epidemiol. 2000;152(10):950-964.

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 3723. doi:10.1158/1538-7445.AM2011-3723
Abstract 3727: Telangiectasias in the skin of patients: Indicator of angiogenic status of tumors?
Authors: Ellberg Carolina , Jernström Helena , Olsson Håkan
Published: June 2012

Background: The angiogenic status of a tumor is related to the prognosis of the patient, and to the possibility of treatment with angiogenesis inhibitors. Patients who express telangiectasias (spiders) in the skin have an increased ability to form vessels, due to, for example, hormonal influences. We hypothesized that tumor tissue to some extent resembles the normal tissue in a patient, which could mean that is if a patient is prone to form spiders in the skin, the same would be true in their tumors. Spiders are associated with hormonal influence, and therefore telangiectasias in patients could be a reflection of hormonal influence and angiogenic status of their tumor.

Materials and methods: A clinical series of 1682 consecutive breast cancer patients diagnosed between 1980 and June 2009 were studied. One physician scored the presence/absence of spiders on the upper part of the body at the time of breast cancer diagnosis. The aim was to further study the association between telangiectasias and classical histopathologic subtypes of breast cancer, HRT-ever use, life time number of menstrual cycles and ever use of oral contraceptive pills (OCP). All analyses were performed using multivariate logistic regression, a confidence interval (CI) of 95% was used.

Results: The occurrence of telangiectasias of the upper thorax, neck and face was positively associated with HRT-ever use (OR = 3.24 (CI 2.02 – 5.19)), OCP-ever use (OR = 2.08 (CI 1.30 – 3.32)) and high number of menstrual cycles, above 550 – the upper quartile OR = 4.57 (CI 1.49-13.95). Weight was also associated with spiders, p = 0.006. Contrary to the other histopathologic subtypes no telangiectasias in the skin were observed in patients with comedo carcinoma, p = 0.0002 (Fisher's exact test). No significant difference in spiders was seen between the other patients with the other subtypes. The results above were adjusted for age at diagnosis and for each studied variable simultaneously.

Conclusion: The occurrence of spiders was associated with several hormonal factors indicating the importance of hormones in the induction of telangiectasias. Our theory that the tendency to form vessels is similar in normal tissue as well as in tumor tissue could explain the necrotic core in comedo carcinoma. It has been hypothesized that the necrotic core is due to the high growth rate of the tumor, but it might be due to a lesser ability to form vessels. Are comedo carcinoma patients a group who benefit less from anti-angiogenic therapy while patients with strong endogenous or exogenous hormonal exposures would benefit more from anti-angiogenic therapy?

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 3727. doi:10.1158/1538-7445.AM2011-3727
Abstract 3728: Night work and breast cancer risk: CECILE study
Authors: Menegaux Florence , Truong Thérèse , Anger Antoinette , Kerbrat Pierre , Arveux Patrick , Lamkarkarch Farida , Guénel Pascal
Published: June 2012

Objectives: Epidemiological studies have suggested that night shift work increases the risk for breast cancer. Disruption of the normal circadian rhythm may explain this association. We investigated the role of night work in the occurrence of breast cancer in a population-based case-control study in France.

Methods: The CECILE study was conducted in two French départements (Côte d'Or, Ille-et-Vilaine) between 2005 and 2008. All female residents in these areas aged 25-75 years who were diagnosed with a breast cancer were eligible to the study. Population controls were frequency-matched by 5-year age group to the cases. A total of 1232 incident breast cancer cases and 1317 controls were included. An in-person interview was conducted using a standardized questionnaire to obtain information on recognized or suspected risk factors for breast cancer, as well as on lifetime occupational history. For each job held for more than 6 months, information on night shift work was elicited. Any work shift where the woman had to work for at least one hour between 11 pm and 5 am was defined as night work. Duration (in years) and frequency (times per week) of night shifts were also obtained. Analyses were conducted using unconditional logistic regression adjusting for age and recognized risk factors for breast cancer.

Results: Thirteen percent of the cases and eleven percent of the controls had ever worked at night (OR=1.29 [1.00-1.64]). The association was more pronounced for women who had overnight shifts (full-time shift work between 11 pm and 5 am: OR=1.37 [1.03-1.83]) than for women who had late evening shifts (ending after 11 pm: OR=1.29 [0.78-2.15]) or early morning shifts (starting before 5 am: OR=1.19 [0.43-3.31]). Women who worked in overnight shifts for 3 or more years had an OR of 1.49 [1.05-1.89], but no further increase of the OR was observed for longer durations. Overnight shift work of 3 nights or less per week during overnight periods, implying frequent changes between night and day schedules, was associated with an OR of 1.59 [1.08-2.33]).

Conclusion: Our results support the hypothesis of a role of overnight work in the occurrence of breast 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 3728. doi:10.1158/1538-7445.AM2011-3728
Abstract 3741: Identification of differentially expressed genes in breast tumors from African American compared with Caucasian women
Authors: Ellsworth Rachel E. , Field Lori A. , Deyarmin Brenda , Love Brad , Hooke Jeffrey , Shriver Craig
Published: June 2012

Background: Breast tumors from African American women (AAW) have less favorable pathological characteristics and higher mortality rates than Caucasian women. While socioeconomic status may influence prognosis, biological factors are also likely to contribute to tumor behavior.

Methods: Patients treated within an equal-access military medical center were matched by age, grade and ER status. Tumors were subjected to laser microdissection and RNA hybridized to HG U133A 2.0 microarrays. Data was analyzed using Partek Genomics Suite using a cutoff of P<0.001, 1.5-fold change. Microarray results were validated by qRT-PCR.

Results: Twenty-six pairs of matched tumor samples were identified. Clinico-pathological factors did not differ significantly for age at diagnosis, tumor size or stage, lymph node, hormone receptor, or HER2 status. Mortality was similar between the two groups. One putative and nineteen known genes were differentially expressed between populations with five genes expressed at higher and 15 genes at lower levels in tumors from AAW; hierarchical clustering classified 23/26 AAW and 26/26 CW correctly. Differential expression was validated (P<0.001) for PSPHL (higher in AAW) and LTF and PSD3 (lower in AAW).

Conclusions: Despite matching of tumors by pathological characteristics and provision of medical care in an equal-access health care system, a molecular signature differentiating AAW from CW tumors was detected, supporting a model in which aggressive tumor phenotypes are driven by molecular differences within the tumors. These differentially expressed genes are involved in immune response, cell cycle and metastasis and thus may contribute to the poor outcome in AAW.

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 3741. doi:10.1158/1538-7445.AM2011-3741
Abstract 3742: Gene-environment Interactions in esophageal adenocarcinoma risk: A case-only analysis
Authors: Zhai Rihong , Zhao Yang , Liu Geoffrey , Ter-Minassian Monica , Wang Zhaoxi , Wu I-Chen , Su Li , Asomaning Kofi , Chen Feng , Kulke Matthew H. , Lin Xihong , Heist Rebecca S. , Wain John C. , Christiani David C.
Published: June 2012

The incidence of esophageal adenocarcinoma (EA) has increased approximately 500% in Western countries over the last four decades. The rapidly increasing incidence and sporadically development feature of EA suggest that gene-environment (G-E) interactions dominate the etiology. However, how G-E contributes to EA carcinogenesis is still poorly understood. We used a case-only approach to test the effect of G-E interactions in the occurrence of EA. 1330 functional and/or tagging SNPs selected from 354 cancer-associated genes were genotyped in 335 Caucasian EA cases. G-E (gastroesophageal reflux symptoms, GERD; BMI; and smoking) interactions were assessed by a two-step approach. First, we applied random forest (RF) to screen for important SNPs that had main effects as well as interaction effects. Second, we used case-only logistic regression (LR) models to estimate the G-E interaction effect, adjusting for covariates and false-discovery rate (FDR). RF analyses identified three sets of SNPs (94 SNPs-GERD, 38 SNPs-smoking, and 44 SNPs-BMI, respectively) that had the highest importance scores and lowest classification error rates. Further case-only LR analysis revealed that multiple interaction markers were significantly associated with EA risk: GERD*rs2237051 of EGF(OR = 1.39, P = 3.50E-07), GERD*rs2440 of XRCC5 (OR = 1,80, P = 0.0008), GERD*rs2237051*rs2440 (OR = 1.85, P = 0.004), smoking*rs10842514 of KRAS (OR = 0.85, P = 0.012), BMI*rs2305742 of IL12RB1 (OR = 0.84, P = 0.003), BMI*rs11568820 of VDR (OR = 0.97, P = 0.0005), BMI*rs11244142 of ABL1 (OR = 1.47, P = 2.09E-05), BMI*rs2800975 of XPA*rs743572 of CYP17A (OR = 3.62, P = 0.0009), and BMI*rs11244142*rs20547 of IL13 (OR = 1.89, P = 0.023). These results indicate that genetic variability may contribute to EA development through interactions with environmental factors. Our data also suggest that G-E interactions may be G- and E-specific in modulating the risk of EA.

Supported by grants: NIH grants CA92824, CA74386, CA90578, and CA119650); Flight Attendant Medical Research Institute (FAMRI) grant 062459_YCSA; the Kevin Jackson Memorial Fund and Alan Brown Chair of Molecular Genomics.

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 3742. doi:10.1158/1538-7445.AM2011-3742
Abstract 3755: Prospective study of genomic hypomethylation of leukocyte DNA and colorectal cancer risk
Authors: Huang Wen-Yi , Su Joseph L. , Hayes Richard B. , Moore Lee E. , Katki Hormuzd A. , Berndt Sonja I. , Weissfeld Joel L. , Yegnasubramanian Srinivasan , Purdue Mark P.
Published: June 2012

Background: Systematic genome-wide (global) reductions of methylated cytosine levels have been observed in most cancer tissues, including colorectal cancer, and are hypothesized to contribute to carcinogenesis through genomic instability and protooncogene activation. Recently, genomic DNA hypomethylation has been measured in peripheral blood leukocytes and associated with colorectal tumors. However, as these studies used post-diagnostic specimens, it is possible that the observed findings reflect disease- and/or treatment-induced effects on leukocyte DNA methylation.

Methods: Using prospectively collected blood specimens and questionnaire data from the Prostate, Lung, Colorectal, and Ovarian (PLCO) Cancer Screening Trial, we studied genomic leukocyte DNA methylation status in relation to subsequent colorectal cancer risk in a nested case-control study of 370 cases and 493 controls with no cancer diagnosis prior to the blood draw. Genomic leukocyte DNA methylation status was determined by an HPLC/Tandem Mass Spectrometry method and expressed as the relative amount of methyl-cytosine to total cytosine residues, or %5-mC. Odds ratios (ORs) and 95% confidence intervals (CIs) were computed using unconditional logistic regression models, adjusting for age, gender, race, time since initial sigmoidoscopy screening, and year of randomization. Inclusion of other suspected colorectal cancer risk factors in the models did not materially change the results.

Results: We observed a relatively narrow range of genomic leukocyte DNA methylation levels among cases and controls, with %5-mC ranging from 3.57 to 4.60 (median = 4.03, standard deviation = 0.19). Categorizing subjects by quartiles of %5-mC content in controls, using the lowest quartile as the reference category (Q1), we observed no trend in risk with adjusted ORs (95% CIs) for Q2, 1.44 (1.01-2.08); Q3, 1.06 (0.75-1.51); and Q4, 1.01 (0.70-1.47) (P-trend = 0.49). Dichotomizing subjects by median of %5-mC in controls with the lower %5-mC group as the reference, we found a borderline-significantly decreased risk for subjects with %5-mC above the median (OR = 0.81, CI = 0.61-1.06), particularly among subjects whose natural folate intake levels were above the median (OR = 0.60, CI = 0.39-0.95; P-interaction = 0.06).

Conclusion: Preliminary findings from our study, to our knowledge the first prospective investigation of DNA methylation and colorectal cancer, show no clear dose-dependent association between genomic leukocyte DNA methylation status and subsequent cancer risk.

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 3755. doi:10.1158/1538-7445.AM2011-3755
Abstract 3759: Meat consumption, ornithine decarboxylase gene polymorphism, and outcomes after colorectal cancer diagnosis
Authors: Lin Bruce S. , Zell Jason A. , Ziogas Argyrios , Anton-Culver Hoda
Published: June 2012

BACKGROUND. Arginine, and meat consumption (i.e., the primary dietary source of arginine) have been implicated in colorectal cancer (CRC) progression via polyamine-dependent processes. Genetic polymorphism in the polyamine-regulatory gene ornithine decarboxylase-1 (ODC1) has been shown to be prognostic for survival after CRC diagnosis. Here we examined the joint effects of meat consumption and ODC1 polymorphism on CRC-specific mortality among CRC cases.

METHODS. The study included 329 incident stage I-III CRC cases from the population-based UC Irvine Gene-Environment Study of Familial CRC (diagnosed 1994-1996 with follow-up through March 2008). Genotyping of the ODC1 (National Center for Biotechnology Information SNP database ID rs2302615) +316 SNP was conducted using oligonucleotide primers designed to amplify a 172-bp fragment containing the polymorphic base at +316 (Applied Biosystems, Foster City, CA). Meat intake (frequency and serving size) was ascertained via questionnaires administered at baseline (cohort entry), where patients were asked to report their usual eating habits during the one year prior to CRC diagnosis. Multivariate CRC-specific survival analyses were stratified by TNM stage, and adjusted for age, gender, race/ethnicity, family history, tumor subsite within the colorectum, histology, treatment with surgery, radiation, chemotherapy, weekly meat consumption, ODC1 genotype, and a term representing the interaction of meat consumption and ODC1 genotype. The primary outcome was to assess the interaction of meat consumption and ODC1 genotype (GG vs. GA/AA) on CRC-specific mortality.

RESULTS. ODC1 genotype distribution was 51% GG, 49% GA/AA). Median meat consumption was 6.95 and 7.40 servings per week in ODC1 GG group and GA/AA group respectively (P =0.31). In the multivariate model, a statistically significant interaction between ODC1 genotype, meat consumption, and CRC-specific mortality was observed, such that increasing levels of meat consumption were associated with higher CRC-specific mortality among ODC1 GA/AA compared to GG cases, adjusted colorectal cancer-specific survival hazards ratio, 12.75; 95% confidence interval, 2.71-60.12; P =0.001 (unadjusted HR 2.29; 95% CI 1.24-4.23; P =0.008). There was a strong interaction between meat consumption and ODC1 genotype, P-int =0.01.

CONCLUSIONS. Among CRC cases with ODC1 GA/AA genotypes, higher levels of meat consumption are associated with poor survival outcomes. These data provide further evidence that polyamine metabolism, and its modulation by specific dietary factors, have relevance to CRC outcomes.

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 3759. doi:10.1158/1538-7445.AM2011-3759
Abstract 377: Multifunctional magnetic nanoparticles for theranostic applications
Authors: Yallapu Murali M. , Othman Shadi F. , Curtis Evan T. , Gupta Brij K. , Jaggi Meena , Chauhan Subhash C.
Published: June 2012

Nanoparticle technology address efficacy of cancer chemotherapy and diagnosis which can be potentially used for image guided drug delivery. Herein, we have developed a multi-layer approach for the synthesis of water-dispersible iron oxide theranostic nanoparticles for drug delivery (therapy) and magnetic resonance imaging (MRI) (diagnostic) applications. For this novel approach, iron oxide core nanoparticles were prepared by precipitation of iron salts in the presence of ammonia and coated with beta-cyclodextrin and pluronic polymer (F127). This formulation (F127250) was highly water dispersible which allowed encapsulation of the anti-cancer drug(s) in beta-cyclodextrin and pluronic polymer for sustained drug release. The F127250 formulation has exhibited superior hyperthermia effects over time under alternating magnetic field compared to pure magnetic nanoparticles (MNP) and beta-cyclodextrin coated nanoparticles (CD200). Additionally, the improved MRI characteristics were also observed for the F127250 formulation in agar gel and in cisplatin resistant ovarian cancer cells (A12780CP) compared to MNP and CD200 formulations. Furthermore, the drug loaded formulation of F127250 exhibited many folds of imaging contrast properties. Curcumin, a cancer preventive and anti-cancer drug, loaded formulation (F127250-CUR) exhibited almost equivalent inhibition effects on A2780CP (ovarian), MDA-MB-231 (breast), and PC3 (prostate) cancer cells even though curcumin release was only 40%. The improved therapeutic effects of this formulation were verified by examining ultra structural changes and molecular effects in cancer cells. Interestingly, F127250-CUR formulation also exhibited good haemo-compatibility. In conclusion, our data suggest that the F127250-CUR formulation can be effectively used for the delivery of anti-cancer drug drugs and cancer imaging and its potential clinical application.

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 377. doi:10.1158/1538-7445.AM2011-377
Abstract 3778: Induction of autophagy and elimination of cancer stem-like cell in glioblastoma multiforme cells by a Magnolia officinalis active component honokiol
Authors: Yao Chih-Jung , Hong Chuang-Ye , Shih Ping-Hsiao , Yeh Chi-Tai , Chen Ruei-Ming , Chuang Shuang-En , Lin Pei-Chun , Yi Tsu-Yi , Lai Gi-Ming
Published: June 2012

The anticancer effects of honokiol, an active component isolated from Chinese traditional herb Magnolia officinalis, had been noticed in recent decade. It had been reported to induce apoptosis, anti-agiogenesis and chemo/radio-sensitization on cancer cells. However, its effects on autophagy induction and cancer stem cell elimination have not yet been investigated. To more evaluate the potential of honokiol in cancer therapy, we explored its effects on these two critical events in glioblastoma multiforme (GBM) DBTRG-05MG cells. The honokiol-induced autophagy was evidenced by the time and dose-dependent increase of LC3-II in treated cells. During this autophagy induction, the phosphorylated AMP-dependent kinase alpha was dose-dependently increased in accompany with the decrease of phosphorylated mTOR protein level. In addition, the phosphorylated p38 was also markedly increased parallel with the elevation of LC3-II. Further investigation on the role of this MAPK p38 pathway in honokiol-induced autophagy is ongoing. In accordance with the growth inhibition and massive cell death induced by high dose of honokiol (50 μM), the phsphorylated Akt and Rb were dramatically suppressed in treated cells. As it is suggested that autophagy is a protective response of cancer cell to environmental stress, an autophagy inhibitor hydroxychloroquine was therefore co-administrated to enhance the effects of honokiol against GBM cells. Significantly, hydroxychloroquine augmented the honokiol-induced cell death, indicating the crucial role of autophagy in honokiol-induced anticancer effects. Moreover, honokiol appeared to have effects on the elimination of cancer stem-like cells. By UV laser-equipped flow cytometer, a small percentage of cancer stem-like side population (SP) cells was detected and sorted from another GBM cell line (GBM 8401). The expressions of stemness genes such as CD133, nestin, Oct4, NOTCH3, IHH and SMO in SP cells were much higher than those in non-SP cells. Treatment with honokiol (5 μM) for 48h decreased the percentage of GBM 8401 SP cells from 1.5% down to 0.2%. These results suggested the potential role of honokiol in GBM treatment.

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 3778. doi:10.1158/1538-7445.AM2011-3778
Abstract 3795: Mitochondrial complex I modulation regulates autophagy and breast cancer progression
Authors: Santidrian Antonio Fernandez , Seo Byoung B. , Matsuno-Yagi Akemi , Yagi Takao , Felding-Habermann Brunhilde
Published: June 2012

Tumor cells express altered metabolic activities often linked to mitochondrial dysfunction. Such mitochondrial defects can inhibit oxidative phosphorylation, change the cellular redox state (NAD+/NADH), increase production of reactive oxygen species (ROS), and cause DNA damage that further supports tumorigenesis and a metastatic phenotype.

Our study identified mitochondrial complex I (NADH dehydrogenase) as a modulator of tumorigenesis and metastatic progression in breast cancer. We used NDI1, the NADH dehydrogenase from yeast, to augment complex I activity in metastatic human breast cancer cells. The NDI1 protein is known to integrate into the inner mitochondrial membrane of mammalian cells, function as NADH dehydrogenase, pass electrons into the respiratory chain, and thereby fully augment intrinsic complex I activity. Thus, NDI1 identifies critical involvement of complex I NADH dehydrogenase activity in cellular functions that are altered upon its expression. We investigated a panel of aggressive human breast cancer cell lines, including MDA-MB 231, and found that augmentation of NADH dehydrogenase activity through NDI1 resulted in an enhanced NAD+/NADH ratio and inhibition of ROS production. Most importantly, NDI1 expression inhibited metastasis and tumor growth in the mammary fad pad of immune deficient mice, seen by non-invasive imaging and histology. The same effects were achieved by treatment with NAD precursors, demonstrating that normalization of the NAD+/NADH ratio is critical for inhibition of breast cancer progression. We found that the underlying mechanism is based on activation of PTEN, inhibition of the AKT/mTOR survival pathway, and stimulation of autophagy. Autophagy is responsible for NDI1-mediated inhibition of breast cancer progression, as knock-down of ATG5 reversed the anti-metastatic effect of NDI1. Our results demonstrate that tumorigenesis and metastasis in breast cancer are critically influenced by mitochondrial complex I activity. We show that augmentation of tumor cell complex I function can interfere with tumor formation and breast cancer spreading through modulating the NAD+/NADH ratio. Thus, normalization of complex I function appears as a promising new target for prevention and treatment of breast progression.

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 3795. doi:10.1158/1538-7445.AM2011-3795
Abstract 3796: Inhibition of mitochondrial respiration synergizes with tyrosine kinase inhibitors to eliminate Bcr-Abl+ leukemia
Authors: Alvarez-Calderon Francesca , Gregory Mark A. , DeGregori James
Published: June 2012

Bcr-Abl kinase inhibitors are the standard therapy for chronic myelogenous leukemia (CML). While effective in controlling disease in chronic phase, these inhibitors often fail to completely eliminate Bcr-Abl+ CML cells and also fail to achieve durable remissions for advanced CML and Bcr-Abl+ acute lymphoblastic leukemias (ALL). Our lab employed a large-scale synthetic lethal shRNA screen to identify genes involved in CML survival upon treatment with the Bcr-Abl kinase inhibitor imatinib (IM) [Cancer Cell. 2010;18(1):74-87]. This screen identified several genes involved in energy utilization including a component of pyruvate dehydrogenase, dihydrolipoyllysine-residue acetyltransferase. This target was identified by two independent shRNAs, each underrepresented over 22 fold in treated cells compared to untreated. CML proliferation depends on Bcr-Abl-driven glycolysis for energy production. Upon IM treatment, I hypothesized that these cells are forced to rely on other sources of energy including the TCA cycle and mitochondrial respiration, and that targeting alternative energy production pathways will enhance IM-mediated elimination of CML cells. Thus, I predicted that inhibition of mitochondrial respiration by knockdown or pharmacologic inhibition would cooperate with tyrosine kinase inhibitors to eliminate Bcr-Abl+ leukemias. My experiments have shown that knocking down pyruvate dehydrogenase in human blast crisis Bcr-Abl+ CML cells (K562) greatly sensitizes cells to IM in vitro. Conversely, supplementing the cell culture media with increasing concentrations of methyl-pyruvate protects K562 cells from IM treatment. Moreover, I find that the mitochondrial ATP-synthase inhibitor Oligomycin A also synergizes with IM to kill CML cells in vitro. In addition, inhibition of mitochondrial respiration with Oligomycin A enhances elimination of Bcr-Abl+ cells by a Bcr-Abl tyrosine kinase inhibitor in vivo and prolongs disease-free survival in a mouse model of Bcr-Abl+ B-ALL. The drug doses of Oligomycin A used are over 100-fold below the LD50, and have no apparent adverse effects on these mice. Targeting mitochondrial metabolism as an adjuvant drug therapy may prevent the survival of residual Bcr-Abl+ CML cells upon tyrosine kinase inhibitor treatment.

*This project is supported in part by pre-doctoral fellowship number F31CA157166 from the National Cancer Institute.

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 3796. doi:10.1158/1538-7445.AM2011-3796
Abstract 3798: P53R2 regulates oxidative phosphorylation complexII and III in keeping integrity of mitochondrial function
Authors: Xue LiJun , Kuo Mei-Ling , Liu Xiyong , Chen Linling , Zhou Bingsen , Yen Yun
Published: June 2012

It has been reported that there is a direct correlation between severe muscle mitochondrial DNA depletion and functionally important mutations in p53R2 in a number of children with mitochondrial depletion syndrome. However, there is no clear evidence how p53R2 involves in the regulation on mitochondrial function. Our current study demonstrated that p53R2 knock-down MCF7 and IMR90 cells displayed low expression level of core proteins in oxidative phosphorylation (OXPHOS) complex II, III, but not complex I, IV or V. Correspondently, activity of complex II and ATP quantity from p53R2-knockdown cells decreased significantly comparing to control cells expressing non-silencing short-hairpin RNA. Moreover, mitochondrial superoxide level increased in p53R2-knockdown cells, which was consistent with the notion that dysfunction of OXPHOS complex II and/or III generated toxic reactive oxygen species in mitochondria. The increased superoxide was not due to SOD2 degradation, since SOD2 expression level was unchanged in IMR90 and MCF7 cells following knockdown of p53R2. These results indicate that down-regulation of p53R2 reduces OXPHOS complex II and III protein expression and correspondent activity. The compromised complex II and III in electron transport chains thus result in premature electron leakage to oxygen to form superoxide in the mitochondria. These observations suggest that p53R2 might protect mitochondria through maintaining integrity of electron transport chains coupling with oxidative phosphorylation.

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 3798. doi:10.1158/1538-7445.AM2011-3798
Abstract 3807: ER-60 increases cell migration and serves as a prognostic marker of cervical cancer
Authors: Liao Chia-Jung , Lin Yang-Hsiang , Lin Kwang-Huei
Published: June 2012

Cervical cancer is the second most common cancer in women worldwide. Human papilloma virus infection is the primary cause of cervical cancer, although it cannot cause cancer by itself. Molecular markers involved in cervical carcinogenesis are needed. A two-dimensional polyacrylamide gel electrophoresis (2-DE) proteomic strategy was used to discover genes with exceptional expression from four paired cervical cancer tissues. Immunoblotting and immunohistochemical analysis with 52 paired specimens and 107 sections were used to confirm the results of 2-DE, and the clinical significance was estimated. The molecular functions of ER-60 were investigated in vitro and in vivo. We identified ER-60 was upregulated in 71.2% of cancerous tissues. ER-60 expression differed significantly between patients with different histological types and differentiation grades (P = 0.013 and 0.045, respectively). Patients with high ER-60 expression had a lower 5-year overall survival (OS) and recurrence-free survival (RFS) rate (P = 0.038 and 0.017, respectively). High ER-60 expression was an independent prognostic factor of poor RFS (P = 0.019). Knockdown of ER-60 expression in HeLa cell decreased cell motility. The lung metastasis was inhibited when introduced ER-60-knockdown cells into severe combined immunodeficiency mice. We observed accumulation of b-catenin and regulation of b-catenin downstream CD44 and S100A4 expression in ER-60-knockdown cell were responsible for this migration inhibition. Taken together, ER-60 overexpression is a potential prognostic factor of poor OS and an independent factor of poor RFS in cervical cancer. ER-60 may be involved in cervical cancer invasion by modulating b-catenin protein stability and signaling.

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 3807. doi:10.1158/1538-7445.AM2011-3807
Abstract 3814: Signet-ring cell gastric carcinoma has different epigenetic signature from poorly differentiated gastric carcinoma
Authors: Kim Hyun Soo , Park Hong Jun , Yang Misuk
Published: June 2012

Background/Aim: Gastric cancer is divided into differentiated intestinal type and undifferentiated diffuse type by the most widely used Lauren's classification, which appear clearly as dissimilar clinical and epidemiological entities. Undifferentiated diffuse type gastric cancer is also classified into poorly differentiated carcinoma and signet-ring cell carcinoma (SRCC). However, recent evidences suggest early SRCC is an initial, differentiated form of diffuse gastric cancer that may evolve into poorly differentiated carcinoma (PDC). To test the hypothesis that molecular epigenetic signature of SRCC is different from that of PDC in early gastric carcinogenesis, we compared epigenetic methylation profile between SRCC and PDC and found unique epigenetic signature of SRCC. Methods: Six genes (GDNF, ROR-a, KLF7, MINT25, E-cadherin (CDH1), and LINE-1) were identified as sensitive molecular markers of early gastric cancer at a previous study. We evaluated the DNA methylation patterns of 4 gene promoters (GDNF, ROR-a, KLF7 and LINE-1(long interpersed nuclear element-1)) by bisulfite-PCR and pyrosequencing in primary tissues from 33 PDCs and 20 SRCCs as a test group. Then, the methylation status of the 6 genes including MINT25 and E-cadherin (CDH1) was next evaluated in endoscopically biopsied gastric cancer tissues from prospectively collected 21 PDCs and 22 SRCCs as a validation group. Results: In the test group, quantitative methylation levels of 3 genes (GDNF, ROR-a, and LINE-1) of SRCCs were significantly different from those of PDCs. MINT25, E-cadherin including the above 4 genes were next evaluated in the validation group. We confirmed the significant different levels of methylations of prospectively collected SRCCs from PDCs. Compared to PDCs, SRCCs showed the lower level of methylations for GDNF (17.7% vs. 31.0%, p = 0.001) and ROR-a (14.4% vs. 28.0%, p = 0.001) genes. Interestingly, however, the methylation level of E-cadherin was higher in SRCCs than that of PDCs (26.8% vs. 20.5%, p = 0.009). Methylaion levels of LINE-1 (63.0% vs. 60.5%), MINT25 (31.1% vs. 34.3%) and KLF7 (10.2% vs. 18.1%) in SRCCs were not significantly different from those of PDCs. Conclusion: Our results identified unique methylation patterns of SRCCs distinguished from PDCs, suggesting different molecular epigenetic signature of SRCC in gastric carcinogenesis.

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 3814. doi:10.1158/1538-7445.AM2011-3814