bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 607: Enhanced inhibition of prostate cancer xenograft tumor growth by combining quercetin and green tea
Authors: Wang Piwen , Heber David , Henning Susanne M.
Published: August 2012

The chemopreventive activity of green tea (GT) and green tea polyphenols (GTPs) in prostate cancer has been demonstrated in preclinical cell culture and animal models. However, extensive methylation of GTPs in vivo to less active methyl-metabolites may be limiting the anti-cancer activity of GT. The objective of the present study was to investigate whether quercetin (Q), a natural inhibitor of catechol-O-methyltransferase (COMT), will increase tissue concentrations of non-methylated GTPs and enhance the inhibition of xenograft tumor growth in severe combined immune deficient (SCID) mice. Androgen-sensitive LAPC-4 prostate cancer cells were injected subcutaneously into SCID mice (n=12 per group). SCID mice were randomly assigned to four groups: control, GT, Q, and GT + Q treatment. The interventions were initiated when tumors reached volume of 10mm3. The final concentration of GTPs in brewed tea administered as drinking water was 0.07% and Q was supplemented in AIN-93G diet at a concentration of 4g/kg diet. There were no differences in food intake, body weight or water consumption during the intervention. Tissue bioavailability of GTPs was increased significantly by 2 to 3-fold in lung, kidney and xenograft tumors and showed a trend to increase by 1.3-fold in liver tissue by GT and Q treatment compared to GT alone. Methylation of epigallocatechin gallate (EGCG), the main polyphenol in green tea, was decreased in tissues from mice in the GT+Q group. After 6 weeks, tumor growth was inhibited by 16% (Q alone), 21% (GT alone), and 45% (GT+Q) compared to the control group. Western blot analysis demonstrated a significant inhibition of methyl transferases such as COMT (31%) and DNA methyltransferase (DNMT) (52%) in tumors from the GT+Q group compared to the control. It has been demonstrated previously that EGCG can act as androgen receptor (AR) antagonist as well as inhibit angiogenesis via inhibition of the vascular endothelial growth factor (VEGF). Together Q+GT inhibited protein expression of AR (35%), prostate-specific antigen (PSA) (39%), and VEGF (25%) more than either GT or Q alone. This study demonstrates an enhanced inhibition of prostate tumor growth when GT and Q are co-administered and this is likely due to inhibition of methylation resulting in enhanced anti-tumor activity of EGCG via effects on angiogenesis, and androgen-mediated tumor growth. This study sets the stage for future human interventions to confirm the increase in bioavailability and chemoprevention using GT in combination with Q. This project was supported by DOD grant W81XWH-10-1-0298 and NIH grant RO3 CA150047-01.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 607. doi:1538-7445.AM2012-607
Abstract 609: Benzyl isothiocyanate inhibits prostate cancer development in the transgenic adenocarcinoma mouse prostate model
Authors: Cho Han Jin , Kim Ji Hee , Park Jung Han Yoon
Published: August 2012

Benzyl isothiocyanate (BITC) is a hydrolysis product of glucotropaeolin, a compound found in cruciferous vegetables, and has also been shown to have anti-tumor properties. In the present study, we attempted to determine whether BITC inhibits the development of prostate cancer using the transgenic adenocarcinoma mouse prostate (TRAMP) model. Male TRAMP mice and their nontransgenic (normal) littermates at 5 wk of age were randomly divided into control and BITC-treatment groups and gavage-fed with 0 (vehicle), 5, or 10 mg/kg of BITC every other day. At the time of sacrifice (24 wk of age), BITC did not affect body weight of normal or transgenic animals. The genitourinary tract weight of TRAMP mice was increased markedly as compared to normal mice, and this increase was suppressed significantly via the oral administration of 5 or 10 mg/kg of BITC. H&E staining of the dorsolateral lobes of the prostate demonstrated that well-differentiated carcinoma (WDC) was a predominant feature in the 24 week old vehicle-fed TRAMP mice, whereas number of lobes with WDC was reduced and the number of lobes with prostatic intraepithelial neoplasia was increased by feeding with 5 or 10 mg/kg of BITC. BITC feeding reduced the number of cells expressing the Ki67 (a proliferation marker) in the prostatic tissue. Additionally, BITC feeding reduced the expression of cyclin E, cyclin D1, cyclin A, and cyclin-dependent kinase (CDK)2. Our in vitro cell culture results revealed that BITC decreased DNA synthesis and CDK2 activity in TRAMP-C2 mouse prostate cancer cells. However, BITC did not affect apoptosis either in transgenic mice in vivo or in TRAMP-C2 cells in vitro. These results indicate that inhibition of cell cycle progression contributes to the inhibition of prostate cancer development in animals treated with BITC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 609. doi:1538-7445.AM2012-609
Abstract 611: Alpha-santalol, a chemopreventive agent induces apoptosis in human prostate cancer cells by causing caspase-3 activation
Authors: Bommareddy Ajay , Rule Brittny , VanWert Adam L. , Santha Sreevidya , Dwivedi Chandradhar
Published: August 2012

The anticancer effects of α-santalol, a major component of sandalwood oil, have been reported against the development of certain cancers such as skin cancer both in vitro and in vivo. The primary objectives of the current study were to investigate the cancer preventive properties of α-santalol on human prostate cancer cells PC-3 (androgen independent and P-53 null) and LNCaP (androgen positive and P-53 wild-type), and determine the possible mechanisms of its action. The effect of α-santalol on cell viability was determined by trypan blue dye exclusion assay. Apoptosis induction was confirmed by analysis of cytoplasmic histone-associated DNA fragmentation using both an apoptotic ELISA kit and a DAPI fluorescence assay. Caspase-3 activity was determined using caspase-3 (active) ELISA kit. PARP cleavage was analyzed using Immunoblotting. α-santalol at 25-75 µM decreased cell viability in both cell lines in a concentration and time dependent manner. Treatment of prostate cancer cells with α-santalol resulted in induction of apoptosis as evidenced by DNA fragmentation and nuclear staining of apoptotic cells by DAPI. α-santalol treatment also resulted in activation of caspase-3 activity and PARP cleavage. The α-santalol-induced apoptotic cell death and activation of caspase-3 was significantly attenuated in the presence of pharmacological inhibitors of caspas-8 and caspase-9. In conclusion, the present study reveals the apoptotic effect of α-santalol in inhibiting the growth of human prostate cancer cells.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 611. doi:1538-7445.AM2012-611
Abstract 616: A cumulative molecular model for the mechanisms of green tea catechins in the chemoprevention of prostate cancer
Authors: Connors Shahnjayla K. , Kumar Nagi B.
Published: August 2012

Background: Prostate cancer (CaP) is the most commonly diagnosed cancer in men. It's long latency, slow progression, and high incidence rates make it ideal for targeting chemoprevention therapies. Studies suggest that green tea may be suitable candidate for CaP chemoprevention. The green tea catechin (GTC) chemopreventive mechanisms against CaP cells, however, are not completely clear. Understanding and refining models of the fundamental molecular pathways by which GTCs modulate prostate carcinogenesis is essential to most appropriately utilize green tea for CaP prevention in clinical settings. Objective: The objective of the study was to review the current literature and develop a model that attempts to recreate the molecular mechanisms and pathways by which GTCs modulate prostate carcinogenesis. Methods: Laboratory studies, clinical studies, and clinical trials focused on GTC chemoprevention of CaP were critically analyzed and reviewed for mechanisms of GTC chemoprevention. The most prevalent mechanisms were combined into a multi-mechanistic model that attempts to recreate the cumulative events that occur as GTCs exert anti-cancer activity on prostate cancer cells and tissues. Results: GTCs exert anti-cancer actions on CaP cells and tissue through six major mechanisms: proteasome inhibition, cell cycle arrest, inhibition of cell proliferation, induction of apoptosis, suppression of carcinogenesis/progression, and inhibition of metastasis. We have proposed a novel model in which GTCs exert chemopreventive effects on CaP though these six major mechanisms that work simultaneously and dependently, largely driven by proteasome inhibition-induced regulation of the NFκB pathway. The cumulative effect of these mechanisms ultimately leads to the inhibition of CaP cell growth, progression, and metastasis. Conclusions: Several mechanisms of GTC chemopreventive activity on CaP cells and tissues have been identified in laboratory and clinical studies. Although GTCs act through distinct cell cycle regulative pathways, the cumulative chemopreventative effect appears to be attributed to their well-coordinated ensemble, rather than a single pathway. Additionally, transitioning to the use of standardized GTC preparations may more accurately reflect human tea consumption, deliver the maximum amount of GTCs, and allow for more generalizable results in epidemiological, laboratory, and clinical studies. We are currently testing and refining our novel cumulative model to fully elucidate the complex mechanism of GTC chemoprevention of prostate cancer in its entirety.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 616. doi:1538-7445.AM2012-616
Abstract 618: Rhodiola rosea L. extract, SHR-5, and metformin exhibit potent activity against bladder carcinogenesis in the UPII-mutant Ha-ras transgenic model.
Authors: Liu Zhongbo , Li Xuesen , Liu Shuman , Xu Xia , Tian Xuejiao , Simoneau Anne R. , Zi Xiaolin
Published: August 2012

The incidence of human urinary bladder cancer increases markedly with age, suggesting a mechanistic connection between aging and bladder carcinogenesis and a potential use of anti-aging agents in bladder cancer chemoprevention. Rhodiola rosea, growing in high altitude or cold regions of the world, has been reported to have anti-aging effects in Drosophila. Metformin, a popular anti-diabetics drug, has also shown anti-aging effect in mice. In addition, both agents have been shown to inhibit the mTOR pathway. Therefore, we examined the effect of SHR-5 and Metformin on bladder carcinogenesis in the UPII-mutant Ha-ras transgenic mouse bladder model. This model mimics human papillary transitional cell carcinoma and exhibits enhanced mTOR activity in the bladder tumors. The homozygous mutant Ha-ras transgenic mice were identified through genotyping using Southern blotting analysis. 0.625% SHR-5 or 0.1% Metformin in drinking water was given to the mice starting at 6 weeks of age and ending at 6 months of age. About 42% of homozygous mutant Ha-ras male transgenic mice which drank normal water died of urinary tract obstruction and hydronephrosis within 6 months of age, while more than 93% and 89% of mice which drank SHR-5 or Metformin containing water survived. The mean bladder weights in male mice drinking SHR-5 or Metformin decreased by 69% and 62%, respectively, compared to control group (Bladder weights for mice drinking normal water versus SHR-5 or Metformin are 0.3382± 0.061versus 0.1278± 0.02 or 0.1053±0.015 gram, P=0.0197 or 0.0082). The mean bladder weights in female mice which drank SHR-5 or Metformin containing water also decreased by about 40% or 39.8 %, respectively, compared to those which drank normal water (Bladder weights for normal water versus SHR-5 or Metformin: 0.07149 ±0.0036 versus 0.04301 ±0.0032 or 0.04295±0.0115 gram, P<0.0001 or =0.0007). Chemstrip tests showed that mice which drank SHR-5 or metformin water developed less hematurine than those which drank normal water. Histological analysis of H&E staining bladder sections from SHR-5 and Metformin treated mice demonstrated more differentiated tumors compared to those from the transgenic mice which drank normal water. Immunohistochemistry staining revealed that SHR-5 and Metformin treatments decreased cell proliferation in the urothelium. Taken together, our results suggest a potential new method for bladder cancer prevention by drinking SHR-5 or Metformin.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 618. doi:1538-7445.AM2012-618
Abstract 639: The effect of grapefruit intake on endogenous serum estrogen levels in postmenopausal women
Authors: Monroe Kristine R. , Stanczyk Frank Z. , Besinque Kathleen , Pike Malcolm C.
Published: August 2012

The inhibitory effect of grapefruit juice on the intestinal cytochrome P450 3A4 system interacts with more than 60% of orally administered drugs leading to elevation of their serum concentrations. While grapefruit has been shown to lead to elevated serum levels of estrogens when hormones are administered orally, there are no published data on the effect of grapefruit intake on endogenous levels. To begin to address that data gap, we conducted a dietary intervention study among healthy postmenopausal volunteers to determine whether endogenous estrogen levels are significantly affected by grapefruit consumption. We tested 4 grapefruit products: whole fresh grapefruit, fresh squeezed juice “not from concentrate,” shelf-stable juice from concentrate, and grapefruit soda. The majority of volunteers were recruited through the Love/Avon Army of Women. The six-week study consisted of a run-in period of three weeks, a two-week period of grapefruit consumption, and a one-week wash-out period. Eight fasting blood samples were collected during the study. An additional 5 samples drawn at 1, 2, 4, 8 and 10 hours after initial grapefruit intake were collected during an acute-phase study. Serum assays for estrone (E1), estradiol (E2), estrone-3- sulfate (E1S), and sex hormone-binding globulin (SHBG) were conducted. Sixty-two women participated in the study, 10 of whom also participated in the acute phase. Only the whole grapefruit had statistically significant effects on endogeneous estrogen levels. The largest increases were seen in the women with the highest BMI. The changes were evident at 24 hours and remained relatively constant over the 2 week period. With whole grapefruit at the average BMI of 26.6 kg/m2, E1S increased overall by 10.6% (p=0.031), and at a BMI of 30 kg/m2 this increased to 16.4% (p=0.003). Approximately double the effects were seen around 10 hours after whole grapefruit consumption. The effects of fresh juice consumption were around 25% of these increases but were not statistically significant. Small increases in E1 were seen with whole grapefruit, but no increase was seen in E2. The findings of the present study suggest an important interaction between grapefruit consumption and serum levels of endogenous E1S. The increase in E1S is difficult to interpret in term of risk of breast cancer as we found only a small increase in E1 and failed to find any increase in E2. More detailed studies are needed to gain an understanding how the increased levels of E1S are not reflected in similar increases in E1 and E2, and whether the increased E1S levels are reflected in an increase in E1 and E2 levels in the breast itself.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 639. doi:1538-7445.AM2012-639
Abstract 642: Body fat distribution and prostate cancer risk: A UK case-control study
Authors: Robinson Fredie , Lophatananon Artitaya , Rahman Aneela A. , Brown Sarah S. , Parker Terence L. , Easton Douglas , Jarai Zsofia K. , Pocock Richard , Dearnaley David , Guy Michelle , Wilkinson Rosemary A. , awyer Emma S , Page Elizabeth , Liu Jo-Fen , Grabowska Anna , Walker David , Hall Amanda L. , Eeles Rosalind A. , Muir Kenneth R. ,   The UK Genetic Prostate Cancer Study Collaborators and British Association of Urological Surgeons
Published: August 2012

Background: Prostate cancer is the most common cancer among males in the UK and in 2007 accounted for almost a quarter of all male cancers diagnosed. Previous studies have shown some associations between body size/obesity and prostate cancer risk. However body fat distribution in terms of perceived “body shape” and its association with prostate cancer risk has never been described. Methods: In the UK data from 1343 cases and 817 controls with age ranged between 38 and 85, with mean age 62.4 (SD 6.2) were collected between 2007 -2009 as part of the UKGPCS Prostate Cancer Study. The data on subjects’ body fat distribution type including “apple” (body fat mostly around the waist and upper trunk), “pear” (body fat around hips), “oval” (body fat generalised on whole body) and “symmetrical” were obtained through self report diagrams in questionnaires after consent from the subjects. Unconditional logistic regression analysis, with adjustment for age, education, ethnic, family history and lifetime body shape size as potential confounders, were carried out to obtain odds ratios and confidence intervals for different types of body fat distribution to assess their possible association with risk of prostate cancer. Results: A “symmetrical” body shape was used as the reference category. Subjects with an “apple” shape were at a significantly 30% reduced risk (adjusted OR 0.70 with 95% CI: 0.55-0.89). Subjects with an “oval” or “pear” shapes did not show any statistically significant association. Conclusion: An “apple” body fat distribution was associated with reduced prostate cancer risk when compared to symmetrical shape.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 642. doi:1538-7445.AM2012-642
Abstract 643: Vitamin D deficiency increases breast cancer risk: A population based case-control study in the Chinese population
Authors: Chen Peizhan , Gu Xiaoli , Liu Yanling , Li Xiaoguang , Li Chenglin , Hu Pingting , Wang Yuan , Ou Zhouluo , Wang Hui
Published: August 2012

Vitamin D plays an important role in metabolism, cellular function and reproduction. Vitamin D has also been suggested to have potential protective effects against many cancers including breast cancer; however, the data from the epidemiological studies have been inconclusive. Our previous meta-analysis have found high dietary vitamin D intake significantly decreased breast cancer risk (RR = 0.91, 95% CI = 0.85-0.97) and women with higher circulating 25(OH)D level showed about 45% (95% CI = 0.38-0.80) decrease in breast cancer risk (Breast Cancer Res Treat. 2010;121(2):469-77). In a population-based case control study with 593 cases and 580 controls from Shanghai area, we have found that 80% of the normal women with severe vitamin D deficiency (less than 20ng/mL), 15.2% were of deficiency status (20 to 30ng/mL). Only 4.8% of women had sufficient vitamin D level (> 30ng/mL). However, in the breast cancer patients, nearly 96.1% were of vitamin D severe deficiency, 3.2% were of vitamin D deficiency and only 0.7% had sufficient levels. Based on a logistic regression analysis, compared to those with the lowest quintile of plasma 25(OH)D level, women with higher level had significant lower breast risk (Q2 vs. Q1, OR = 0.45, 95% CI = 0.33-0.63; Q3 vs. Q1, OR = 0.25, 95% CI = 0.17-0.36; Q4 vs.Q1, OR = 0.09, 95% CI = 0.06-0.15) showing a significant dose-effect relationship (p-trend < 0.001) after adjusted for the co-variants. We also found a seasonal distribution of vitamin D. Significantly lower vitamin D levels were found for the cases and controls in spring and summer (p < 0.01). In the controls the mean level of vitamin D was higher in the summer (17.76ng/ml) and autumn (16.25ng/ml), and lower in the spring (15.67ng/ml) and winter (10.75ng/ml). However, in the breast cancer patients, the plasma level of vitamin D was highest in the autumn (14.38ng/ml) and lowest in the spring (10.16ng/ml). These results provide a strong evidence that vitamin D may have a chemo-preventive effect against breast cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 643. doi:1538-7445.AM2012-643
Abstract 647: Use of statin and risk of prostate cancer recurrence among patients who received radical prostatectomy
Authors: Chao Chun R. , Xu Lanfang , Wallner Lauren P. , Porter Kimberly P. , Jacobsen Steven , Williams Stephen G.
Published: August 2012

Background: HMG-CoA reductase inhibitors (“statins”) have shown broad-spectrum anti-cancer properties in laboratory studies. In epidemiologic studies, statin use has been linked to reduced risk of advanced prostate cancer (PCa). We investigated whether statin use is associated with reduced risk of recurrence in PCa patients who received radical prostatectomy. Methods: All men with incident PCa diagnosed between 2004-2005 who subsequently underwent radical prostatectomy in Kaiser Permanente Southern California (KPSC) health plan were identified using KPSC's accredited cancer registry. Exclusion criteria included <24 months prior KPSC membership, use of neoadjuvant therapy, and stage 4 disease. Subjects were followed for 5 years after prostatectomy for: (1) Biochemical recurrence, defined as a single PSA measurement >0.2 ng/ml; and (2) clinical disease progression, defined as diagnosis of metastatic disease (identified using ICD-9 codes) or PCa-related death (i.e., PCa as primary cause of death). Information on statin use, demographics, co-morbid conditions, pathoclinical prognostic factors, e.g., stage and Gleason score, and outcomes were obtained from cancer registry and electronic medical records. The effects of statin use prior to and after prostatectomy were both examined using bivariate and multivariable Cox models, adjusting for known prognostic factors. For post-operative statin exposure, a time-dependent Cox model was used. Results: A total of 1192 men were included; 38% had pre-operative and 55% had post-operative statin use. Neither pre- nor post-operative statin use was associated with risk of any outcome, as there was no evidence of a clear dose-response relationship (Table). Conclusion: These data suggest that despite statin's protective effect in developing PCa, statin use may not prevent PCa progression.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 647. doi:1538-7445.AM2012-647
Abstract 661: Physical activity, sitting time, and postmenopausal breast cancer risk by histopathologic characteristics in a large US cohort
Authors: Patel Alpa V. , Hildebrand Janet S. , Gaudet Mia M. , Teras Lauren R. , Campbell Peter T. , Gapstur Susan M.
Published: August 2012

There is convincing evidence that physical activity is associated with lower risk of postmenopausal breast cancer, likely through its impact on circulating sex hormones, improved insulin sensitivity, and/or weight control. However, the relationship between time spent sitting and breast cancer is not well-defined. Furthermore, it is unclear whether the relationship of physical activity or sitting time with postmenopausal breast cancer varies by histopathologic characteristics of the tumor (defined by stage, histology, or estrogen/progesterone receptor (ER/PR) status), use of postmenopausal hormones (PMH), or body mass index (BMI). Therefore, we examined recreational physical activity and leisure-time spent sitting in relation to breast cancer risk among 73,608 postmenopausal women who were cancer-free and enrolled in the American Cancer Society Cancer Prevention Study II (CPS-II) Nutrition Cohort, a prospective study of cancer incidence and mortality that began in 1992. From 1992-2007, 4,622 incident breast cancer cases were identified. Cox proportional hazards regression modeling was used to compute rate ratios (RR) and 95% confidence intervals (CI) while adjusting for potential confounders including age, race, education, BMI, PMH use, and other breast cancer risk factors. Women who engaged in the highest amount of physical activity (>42 MET-hours/week) had 22% lower risk of breast cancer compared to women with the least activity (0-<7 MET-hours/week) (RR 0.78, 95% CI 0.63-0.95; trend p=0.09). Women who walked an hour per day were 15% less likely to develop breast cancer during follow-up compared to women who walked fewer than 3 hours per week (95% CI 0.76-0.95). Sitting time was not associated with breast cancer risk (RR=0.98, 95% CI 0.89-1.09 for 6+ vs <3 hours/day spent sitting). When examining associations by stage, the association between physical activity and breast cancer risk was only observed for localized breast cancer (RR=0.67, 95% CI 0.51-0.89; trend p=0.03), and not for in situ or regional/distant stage. The association did not differ by histology or ER/PR status. The association between physical activity and risk was not modified by BMI (interaction p=0.73) or by PMH (interaction p=0.74), although the lower RR associated with the highest level of physical activity was noted only in non-users of PMH. These findings provide further evidence that physical activity is associated with lower risk of postmenopausal breast cancer and that the benefits of physical activity do not appear to be limited to any specific subgroup of women.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 661. doi:1538-7445.AM2012-661
Abstract 674: A pilot study of physical activity and breast cancer risk in Africa
Authors: Hou Ningqi , Gakwaya Antony , Ndom Paul , Ojengbede Oladosu , Olopade Olufunmilayo I. , Huo Dezheng
Published: August 2012

Background: As a modifiable risk factor, physical activity (PA) has been linked to breast cancer, and there is cumulative evidence for a beneficial effect of PA on breast cancer risk. However, most of the studies were conducted in developed countries, with little knowledge on PA level and its relation to breast cancer risk among indigenous African populations, where breast cancer may have unique etiology. Methods: We used preliminary data from the African Breast Cancer Study (ABCS), a recently launched, multicenter case-control study, conducted in Cameroon, Nigeria, and Uganda in Africa. After consulting with native investigators in Africa, we developed a physical activity questionnaire to assess the type, frequency, intensity, and duration of various activities. Based on the questionnaire, we calculated sub-scores of housework PA, occupational PA, and leisure PA, as well as a total PA score estimating daily energy expenditure (MET-hours/day). Multivariate linear regression was used to examine the relationship between the total score and sub-scores of PA and case-control status, age, and study sites. Results: To date, 98 cases and 120 controls have been enrolled and the PA questionnaires were administered. The mean age was 48.6 years, ranging from 19-84 years. The overall PA score averaged at 29.6 MET-hours/day, mainly composed by housework PA (an average of 20.5 MET-hours/day) and occupational PA (an average of 16.2 MET-hours/day), with relatively low level of leisure PA (an average of 1.3 MET-hours/day). PA scores were the highest in the Cameroon study site, and the lowest in the Nigerian study site. PA score varied by age, but there was no clear trend. In the multivariable regression model, case-control status was significantly associated with the total PA score: cases were estimated to have 14.0 MET-hours/day lower PA compared to controls, after adjusting for age and study site (p<0.05). Conclusion: African women's physical activity mainly consists of housework activities and work-related activities, with little activities conducted for recreational purposes. The preliminary data show physical activity may be significantly associated with reduced breast cancer risk, but further investigation and a larger sample size are to be followed.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 674. doi:1538-7445.AM2012-674
Abstract 690: Discovery and preliminary validation of novel early detection biomarkers for triple-negative breast cancer using preclinical plasma samples from the Women's Health Initiative observational study
Authors: Li Christopher I. , Mirus Justin , Zhang Yuzheng , McIntosh Martin W. , Lampe Paul D.
Published: August 2012

Background: One approach to develop new tools for detecting cancer early is through the identification and validation of blood-based cancer specific biomarkers. In applying this approach to breast cancer one potential challenge is the disease's considerable heterogeneity. Triple-negative (TN) breast cancer is a particularly lethal breast cancer subtype that is known to have a unique biology. Consequently, there is a clinical need to discover early detection biomarkers for TN disease, and such biomarkers may be more readily identifiable given the more aggressive nature and growth patterns of these tumors. Objective: To discover and initially validate novel TN breast cancer specific early detection biomarkers using preclinical plasma samples. Methods: We conducted a nested case-control study of TN breast cancer within the Women's Health Initiative (WHI) Observational Study (OS) consisting of 28 TN cases whose blood was drawn 17 months prior to their breast cancer diagnosis and an equal number of matched controls. The 28 matched sets were equally divided into a training set, used for discovery, and an independent test set, used for validation, and interrogated using a customized antibody microarray. Results: In the training set, of the 915 antibodies assessed, 55 (6.0%) were statistically significantly different between cases and controls at p<0.05 and in the test set 128 (14.0%) were statistically different. Of the 55 candidates identified in the training set, 15 were validated in the test set at p<0.05. Of these 15 candidates, 13 had higher levels in cases compared to controls. The candidates with the strongest signals included myosin-5A (log2 ratio=1.26), arfaptin-2 (log2 ratio=1.02), integrin-beta1 (log2 ratio=0.99), phosphoserine aminotransferase (log2 ratio=0.94), beta-catenin (log2 ratio=0.92), mast/stem cell growth factor receptor (log2 ratio=0.90), and IGFBP-2 (log2 ratio=−1.18). Conclusions: Several unique, novel candidate early detection biomarkers specific to TN breast cancer were discovered and initially validated in preclinical samples. These findings warrant validation in additional sets of TN cases and controls to further evaluate their potential clinical significance.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 690. doi:1538-7445.AM2012-690
Abstract 691: Estrogen receptor mRNA level in breast cancer predicts response to tamoxifen
Authors: Bordeaux Jennifer , Cheng Huan , Welsh Allison , Haffty Bruce , Lannin Donald , Wu Xingyong , Su Nan , Ma Xiao-Jun , Luo Yuling , Rimm David
Published: August 2012

PURPOSE: Quantification of mRNA has historically been done by reverse transcription polymerase chain reaction (RT-PCR). Recently, a robust method of detection of mRNA utilizing in situ hybridization has been described that is linear and shows high specificity with low background. Here we describe the use of the AQUA method of quantitative immunofluorescence (QIF) for measuring mRNA in situ using ESR1 (the estrogen receptor alpha gene) in breast cancer to determine its predictive value compared to Estrogen Receptor ≤ (ER) protein. METHODS: Messenger RNA for ER (ESR1) and Ubiquitin C (UbC) were visualized using RNAscope probes and levels were quantified by quantitative in situ hybridization (qISH) on two Yale breast cancer cohorts on tissue microarrays. ESR1 levels were compared to ER protein levels measured by QIF using the SP1 antibody. RESULTS: ESR1 mRNA is reproducibly and specifically measurable by qISH on tissue collected from 1993 or later. ESR1 levels were correlated to ER protein levels in a non-linear manner on two Yale cohorts. High levels of ESR1 were found to be predictive of response to tamoxifin. CONCLUSION: Quantification of mRNA using qISH may allow assessment of large cohorts with minimal formalin fixed, paraffin embedded tissue. Exploratory data using this method suggests that measurement of ESR1 mRNA levels may be predictive of response to endocrine therapy in a manner that is different from the predictive value of ER. Further studies are underway using tissue microarrays from other institutions to determine how tissue age affects this assay.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 691. doi:1538-7445.AM2012-691
Abstract 693: Development of a protein-based functional assay for the determination of BRCA1 carrier status
Authors: Bathurst Lauren E. , Uy Paolo , Feilotter Harriet , Davey Scott
Published: August 2012

While most breast cancers are sporadic, 5-10% are hereditary and attributable to the presence of a mutation in the breast cancer associated gene 1 (BRCA1). The BRCA1 gene encodes a tumour suppressor with several well described roles in maintaining genomic integrity, such as cell cycle checkpoint control and DNA repair. BRCA1 is also known to play a role in the differentiation of breast epithelial cells. Given the high risk of cancer development associated with BRCA1 mutation carriers, it is important that they be identified early and accurately. Previous work in our lab developed a novel functional assay to predict BRCA1 status based on gene expression profiles. That study analyzed EBV-transformed lymphocyte cell lines (LCLs) of BRCA1 mutation carriers (BRCA1+/−) and controls (BRCA1+/+) and found that lymphocytes from BRCA1 carriers could be distinguished with high fidelity (∼90% accuracy) from individuals with two wild type copies of the gene. Interestingly, it was found that many of the genes used to distinguish BRCA1 mutation carriers were also markers of differentiation in blood cells. We are now using this information to develop a flow cytometric protein-based biomarker assay. To do this, our transcriptome microarray data was analyzed to identify genes with strong edictability of BRCA1 status, including genes with known roles in lymphocyte differentiation such as TBX21 and CXCR3. These genes were then used to model the assay in-silico using a training set of data comprised of 53 LCLs. The model was designed by selecting genes both up- and down-regulated in BRCA1 carriers, placing them in pairs and looking at the ratio of their expression values. This model was subsequently validated using a test data set comprising 16 LCLs. Overall accuracy of the model was 94%, correctly predicting 54/59 LCLs. Based on in-silico modelling, we chose six antibodies to develop the assay. To identify other candidate proteins, LCL lysates were analyzed by mass spectrometry (MS). The results identified several other candidate proteins for assay development including IgG1, IgD, BCR and MX1. Interestingly, as was the case in our transcriptome analysis, these MS identified proteins also suggested that BRCA1 mutation carriers displayed defects in cellular differentiation. This was indicated by the higher expression of surface markers in responding to less differentiated B-lymphocytes in comparison with controls. Ultimately, the assay will be applied to fresh patient blood samples with the expectation that it will become an assay that can be implemented in a clinical setting to provide accurate identification of BRCA1 mutation carriers and aid in disease management.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 693. doi:1538-7445.AM2012-693
Abstract 698: Overexpression of the aryl hydrocarbon receptor correlates with high tumor grade in human breast invasive carcinomas
Authors: Eltom Sakina E. , Prins Petra , Olivares Maria G. , Ballard Billy R. , Johnson Kimberly N. , Granja-Ingram Nara D. , Agboto Vincent K. , Ding Tan
Published: August 2012

The aryl hydrocarbon receptor (AhR) is a ligand-activated basic helix-loop-helix transcription factor which binds environmental poly aromatic hydrocarbons (PAH) and mediates their carcinogenic effects. Recent reports, including data from our laboratory implicate AhR in breast cancer development and progression independent of the receptor occupancy by PAH. In this study we investigated the potential of AhR as a stage specific marker of breast cancer. We examined the expression of AhR by immunohistochemistry in tissue microarrays (TMA) containing 192 specimens of clinically defined three stages of invasive breast cancer: node-negative (NN), node-positive (NP) and metastatic (Met) carcinoma. The TMA were obtained from the National Cancer Institute Cooperative Breast Cancer Tissue Resource [NCI-CBCTR] and were stained for AhR using high affinity antibody. The AhR staining was then scored by three evaluators, including two pathologists who were blinded to the study. Statistical analysis showed a significant correlation between the AhR expression and the carcinoma case type (NN, NP or Met) (p-value of ANOVA is < 0.0003), and although there is a significant difference of AhR expression among carcinoma case type of white women (p< 0.0017) there is no significant difference for black women (p<0.0968). There is also a strong positive correlation between the receptor expression and the tumor grade (p <0.0001). While there is no correlation with the status of estrogen or progesterone receptors (p= 0.1643 and 0.1884, respectively), there is strong correlation of high AhR expression and invasive breast carcinoma in women over 50 years old (p<0.0043). In conclusion, our findings identify the AhR as a new predictive clinical marker for metastatic breast cancer and a unique target for the design of novel selective inhibitors for therapeutic intervention of metastatic breast cancer. More importantly, the AhR overexpression identifies a subset of patients who could benefit from therapy targeting this receptor.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 698. doi:1538-7445.AM2012-698
Abstract 701: TUMOR lymphocyte infiltration, expression of CD4, CD8, FOXP3, CD34 molecules and their relationship with tumor evolution after primary treatment in breast cancer patients
Authors: Rico María J. , Rozados Viviana R. , Herrera Cintia , Alasino Carlos M. , Roig Laura M. Vargas , Gago Francisco E. , Tello Olga , Nocito Ana L. , Roggero Eduardo , Scharovsky Olga G.
Published: August 2012

Considering that, approximately, only half of breast cancer patients respond to present therapies, a better knowledge of breast cancer biology, one of the most common neoplasms in women, will enable to design better therapeutic approaches. It has been proposed that different tumor characteristics like inflammatory infiltrate intensity, number of T regulatory (Tregs) cells and density of tumor vasculature could be useful to anticipate the response to therapy. Our aim was: a) to analyze the lymphocytic infiltrate and the expression of CD4, CD8, CD34 (endothelial marker) and Foxp3 (Treg marker) in primary tumors of breast cancer patients, and b) to relate them with clinical evolution (disease free: DF or relapsed: R) after 5 years from the primary treatment (surgery). There were analyzed archive samples of ductal mammary tumors (n=27), mostly in stages I and II. The expression of CD4 and CD8 in intra- and peri-tumoral lymphocytes was determined by immunohistochemistry (IHC) and confirmed by confocal microcopy; also the expression of Foxp3 in lymphocytes and tumor cells and CD34 in endothelial cells was evaluated by IHC. The lymphocytic infiltrate was assessed in histological sections stained with hematoxylin-eosin. The quantification was done in 20 high magnification fields and a score ranging from 0 (null) to 6 (high) was utilized. In DF patients (n=22) the intratumoral lymphocytic infiltrate (median [range]: 3 [0-6]) was more intense than that of R patients (n=5), (2 [0-2]), (P=0.05). For CD4, CD8, CD34 and Foxp3 molecules no differences between groups of patients were observed, being their expression highly variable: the score for CD34 varied from 1 to 6 and for the other three molecules from 0 to 6, in either group. In conclusion, the presence of an intense intratumoral lymphocytic infiltrate could be a prognostic marker of good response to treatment, at least in the first 5 years after surgery. These results warrant the study of higher number of tumor samples, since its confirmation could give support for its use in the clinical setting.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 701. doi:1538-7445.AM2012-701
Abstract 702: Molecular responses to conductive interstitial thermal therapy of murine 4T1 breast carcinoma in surviving tumors, bone marrow and serum.
Authors: Przybyla Beata D. , Vishal Sagar
Published: August 2012

Background: Ablation of solid tumor with a laser probe (CITT therapy) causes coagulative necrosis in central zone of tumor. The exposure to temperature gradient and immune responses to this treatment may affect homeostasis of surviving cells in surrounding area as well as induce systemic changes. The purpose of this study was 1/ to perform quantitative analyses of 29 transcripts of selected genes in tumor remaining after ablation and in bone marrow 2/ to evaluate changes in 16 cytokines in serum 3/ to compare levels of heat shock proteins in surviving tumors and in control tumors. Methods: Real-time RT-PCR and “delta delta” method was used to determine changes in transcripts in remaining tumors after treatment relative to control tumors. Cytokines in serum were tested using multiplex assay consisting of 16 murine cytokines. Expression of Hsp27 and Hsp70 proteins were determined with Western Blot. Results: 72 hours after thermal ablation of 4T1 tumors the following changes were found in tumor, in bone marrow and in serum of treated mice: 1/Out of 29 quantified transcripts, the level of four transcripts (Csf3 Lifr, Serpin1 and Vcam1) decreased significantly in remaining tumor. Majority of quantified transcripts had lower level in ablated than in control tumors. In bone marrow (BM) an opposite trend was observed. Levels of majority of quantified transcripts in BM increased after ablation as compared to control mice. The transcripts of genes: Cxcl12, Sele, Fgf2, Lifr, Mmp9, Timp3 were significantly higher in BM of mice after tumor ablation. 2/ Out of 16 chemokines tested, 4 chemokines (Il-5, Il-6, Ccl2 (MCP-1) and Ccl5 (RANTES) were detected in serum. There was a consistent increase of Il-5 in serum from ablated animals. 3/ Levels of Hsp27 and Hsp70 proteins were higher in remaining tumor as compared with control tumors. Conclusion The decrease of transcripts in remaining tumor of genes involved in adhesion, extracellular matrix degradation and tumor viability may inhibit re-growth of thermally ablated tumors. The increase of transcripts in bone marrow of treated mice indicates alteration of homeostasis of vascular niche and changes in function of mesenchymal stem cells. The increase of Hsp27 may inhibit apoptosis in remaining tumor. Up-regulation of Interleukin 5 suggests that this cytokine may play role in stimulation of anti-tumor immunity, as observed by other researchers. Further study is needed to determine whether the transcript reported here may be used as markers predicting long term effects of thermal ablation of solid tumors.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 702. doi:1538-7445.AM2012-702