bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract P4-05-11: A proteasomal signature of breast cancer cells
Authors: Gaczynska M , Mueller B , Rodriguez KA , Osmulski PA 
Published: August 2014

Abstract
The multisubunit, multifunctional protease called proteasome as a part of the essential proteasome-ubiquitin pathway constitutes an attractive target for anti-cancer drugs. A proteasome inhibitor bortezomib is successful in blood cancers treatment, however trials with breast cancers yield disappointingly inconsistent results, likely stemming from the cancers’ diversity. Since bortezomib preferentially induces an apoptosis in cancerous cells, it is likely that differences in the proteasome pathway between the control and cancerous cells, and also between distinct cancers, contribute to this effect. Here we compared components of proteasome assemblies and their peptidase activities in seven breast cancer and two control cell lines in an attempt to find a common proteasomal signature of breast cancer. Defining a common proteolytic denominator for breast cancer cells should facilitate a rational drug design based on proteasomal inhibition. For this purpose, we analyzed cytosolic, nuclear and microsomal extracts from human cultured breast cells: control (MCF10A, Hs 578 Bst) and derived from distinct types of breast cancer: luminal A MCF7 and T47D, triple negative basal A MDA-MB-468, and triple negative basal B MDA-MB-231, Hs 578 T and BT-549. We found a high content of total proteasome in a microsomal fraction with a relatively prominent contribution from immuno proteasomes in all cancer cell lines. These proteasomes are believed to be localized on the outside of endoplasmic reticulum membrane and presumably take part in the endoplasmic reticulum - associated degradation system. Markedly, the pattern of peptidase activities of proteasome and content of subunits of its exchangeable sub-assemblies were specific for each fraction and discriminated between cancerous and control cells. Statistical analysis identified two clusters formed by the cell lines in the microsomal and nuclear fractions and a distinct clustering of the cytosols. Furthermore, three groups of variables, which tend to always cluster together were identified: the 19S regulatory particle subunits, immunoproteasome subunits and peptidolytic activities. Variables that the best quantitatively discriminate between the cell lines at a given level of total proteasome were immunoproteasome subunits and 19S cap subunit Rpt5. Additionally, the housekeeping subunit beta5 in the cytosol, the PA28 activator gamma subunit in the microsomal fraction and chymotrypsin like activity in the nuclear fraction qualitatively were the best markers aiding classification of the cell lines. We speculate that the distinct properties of proteasomes in the microsomal and nuclear fractions may contribute to specific response of the cancer cell lines to the treatment with proteasomal inhibitors. The study shows that a comprehensive analysis of the proteasome system in the established breast cancer cell lines may help to guide diagnostics and therapy design.
Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-05-11.
Abstract P4-06-13: Discovering serum biomarkers for predicting accumulation of nanotherapeutics to different tumor and organ microenvironments
Authors: Yokoi K , Tanei T , Godin B , van de Ven AL , Alexander J , Saito Y , Ferrari M 
Published: August 2014

Abstract
Surrogate biomarkers in the circulation for the enhanced permeation and retention (EPR) effect in different tumor and organ microenvironment shall aid in selecting patients who are likely to accumulate higher amount of nanotherapeutics to tumors and thus show better response to the therapeutics. In this study, 4T1 and 3LL murine cancer cell lines, known to have similar sensitivity to pegylated liposomal doxorubicin (PLD) in vitro, were shown to accumulate significantly different quantities of PLD in vivo. PLD accumulation correlated with tumor-specific differences in therapeutic efficacy and vascular permeability, which was modulated by the extent of coverage of tumor-associated endothelial cells by basement membrane. Matrix metalloproteinase (MMP)-9 and its endogenous inhibitor, tissue inhibitor of metalloproteinase (TIMP)-1, played a pivotal role in these phenomena. Differences in vascular permeability and PLD accumulation were also organ-specific and significantly correlated with the relative ratio of MMP-9 and TIMP-1 in the systemic circulation of the tumor-bearing mice. Our findings support the further development of MMP-9 and TIMP-1 as paired serum biomarkers for predicting accumulation of PLD to tumor and the application of these biomarkers could be expanded to other types of nanotherapeutics utilizing the EPR effect for the personalization of cancer therapy.
Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-06-13.
Abstract P4-07-14: The effects of calorie restriction on microRNA expression profiles and DMBA-induced mammary tumorigenesis in a model of luminal A breast cancer
Authors: Devlin KL , Sanford T , Mambo E , Hursting SD 
Published: August 2014

Abstract
MicroRNA (miR) are small, post-transcriptional regulators that play an integral role in maintenance of cellular functions and whose dyregulation has been shown to promote many types of cancer, including breast cancer. While it has been shown that microRNA are dyregulated through the stages of breast cancer development, it was our goal to define the microRNA expression profiles of the individual stages in a model of luminal A breast cancer in order to identify which are specifically associated with progression. In addition, we aim to establish the effects of calorie restriction, which is known to inhibit breast cancer progression, on relevant microRNA. To address these questions, 100 female Sprague Dawley rats were administered either dimethylbenz(a)anthracene or vehicle control at 50 days of age, then randomized to receive either control (AIN-76A) diet ad-libitum (n = 40) or a 30% CR diet regiment (n = 60). Resultant mammary tumors were allowed to develop for 12 weeks. Calorie restriction significantly increased survival to study endpoint relative to control diet (75% vs 35%, respectively) (p = 0.0047). Furthermore, of the animals that developed tumors, CR significantly decreased median tumor area by 56% compared to control diet (109.4 mm2 vs 250.9 mm2, respectively) (p = 0.0286). To analyze the impact of microRNA modulation on tumor progression, mammary tumors from control animals were microdissected in a stage-dependent fashion, with areas of normal epithelium, intraductal proliferation, ductal carcinoma in-situ, and invasive ductal carcinoma collected and analyzed separately. We have identified 10 microRNAs that are associated with cancer progression in this model (miR-10a, -10b, -21, -92b, -124, -125b, -126, -145, -200a, -658). We are currently analyzing which of these microRNAs are CR-responsive and how their modulation affects tumorigenic indices. The results obtained will provide insights into the mechanisms of breast cancer progression and how calorie restriction inhibits progression through microRNA modulation.
Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-07-14.
Abstract P4-07-19: MicroRNA miR-24 enhances tumor angiogenesis and metastasis in breast cancer
Authors: Xu Z , Wang X , Fang L 
Published: August 2014

Abstract
MicroRNAs (miRNA) are non-coding single-stranded RNA molecules, which are produced from endogenous transcripts of short stem-loop structures. Through binding to the 3’-untranslated region (3’UTR) of different target messenger RNAs (mRNAs), microRNAs can repress the translation of the target mRNAs. Recent studies have shown that miR-24 can inhibit apoptosis of cancer cells and cardiomyocytes, and can accelerate cell proliferation. Up-regulation of miR-24 has been observed in oral carcinoma and is one of the most abundant miRNAs in cervical cancer. However, the role of miR-24 in breast cancer cell angiogenesis and metastasis is not clear.
We analyzed microRNA miR-24 levels in patients with breast carcinoma and found that miR-24 was higher in breast carcinoma samples than in breast benign tissues. We generated constructs expressing miR-24 and studied its functions using both in vitro and in vivo techniques. We found that the ectopic expression of miR-24 promoted breast cancer angiogenesis and metastasis.
Cell migration assays indicated that expression of miR-24 promoted migration of MT-1 cells and 4T1 cells. We examined proliferation of MT-1 cells and 4T1 cells transfected with miR-24 or the control vector. The experiments showed that the miR-24 transfected cells had high rates of proliferation than the vector-transfected cells. In vivo experiments indicated that the expression of miR-24 enhanced tumor growth, metastasis to lung tissues, and decreased overall mouse survival. Tumor sections were also probed with anti-CD34 antibody to examine angiogenesis. We found that miR-24 expression increased tumor-associated vascularization. We further performed tumor metastatic assays. DNA was isolated from lung tissues and subjected to PCR to indicate metastasis of the cancer cells. Typical metastatic lesions in the lungs are shown.
In the miR-24 expressing cells and tumors, expression of the phosphatases PTPN9 and PTPRF were repressed. We confirmed that miR-24 could directly target both PTPN9 and PTPRF. Consistent with this, we found that the levels of PTPN9 and PTPRF were lower in the patients with metastatic breast carcinoma. Ectopic expression of PTPN9 and PTPRF decreased cell migration, and tumor metastasis.
Our study demonstrates that miR-24 is highly expressed in human breast carcinoma. This finding suggests a role of miR-24 in breast cancer development/progression. Our results suggest that miR-24 plays a key role in breast cancer angiogenesis and metastasis. miR-24 could potentially be a target for cancer intervention.
Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-07-19.
Abstract P4-09-02: Omega-3 ethyl esters suppress breast cancer growth by modulating inflammatory signaling
Authors: Chen CH , Quach D , Bowers L , Fabian C , Hursting S , deGraffenried L 
Published: August 2014

Abstract
Introduction: Inflammation has been identified as a key contributor to breast cancer development. Both clinical and preclinical studies confirm a role for inflammatory mediators such as cytokines, interleukins, embedded immune cells and prostaglandins in promoting development of breast cancer. Elevated levels/activity of cyclooxygenase-2 (COX-2) and interleukin-6 (IL-6) are correlated with a more aggressive disease. Omega-3 fatty acid (n-3) intake is correlated with an inverse risk for breast cancer development and improvement in prognostic markers. One key target of omega-3 fatty acids is the COX-2 enzyme. We hypothesize that one mechanism by which omega-3 fatty acids suppress breast cancer progression is through inhibition of inflammatory signaling.
Methods: The impact of omega-3 ethyl esters (n-3 EE), a component of some omega-3 supplements, on the viability of MCF-7 breast cancer cells grown in a pro-inflammatory environment was assessed by MTT analysis and on proliferation by cell counting. Supplementation of the growth media with IL-6 (10ng/mL) was used to simulate a pro-inflammatory environment. Changes in expression levels of key components of inflammatory pathways were assessed by Western blot analyses and quantitative PCR. Prostaglandin E2 (PGE2) levels were measured using ELISA assays.
Results: A significant suppression in IL-6-induced proliferation was observed when cells were exposed to physiological concentrations (20 uM) of n-3 EE for 96 hrs. Molecular analyses suggest that the suppression of the NF-kB/COX-2/PGE2 signaling axis was important for mediating this effect. These results are consistent with other studies using specific COX-2 inhibitors.
Conclusions: With their potent anti-inflammatory activity, n-3 EE may prove useful in reducing malignancy of breast cancer and also slow development of new breast cancers. Importantly, they appear to have none of the toxicities associated with pharmaceutical COX inhibitors (NSAIDs). Future studies are planned to incorporate nutraceutical compounds to standard therapy to improve efficacy and reduce associated side effects.
Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-09-02.
Abstract P4-09-06: No inverse association between breast cancer risk and n-3 unsaturated fatty acids in erythrocytes among Japanese women
Authors: Koriyama C , Kijima Y , Natsugoe S , Akiba S 
Published: August 2014

Abstract
Dietary intake of fish, a major source of docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), has been proposed to reduce the cancer risk. In contrast to the animal studies, their protective effects for breast cancer are equivocal in epidemiological studies. In a case-control study, we examined the association between the ratio of DHA (EPA) to aracidonic acid (AA) in erythrocytes and breast cancer risk in Kagoshima, Japan. All subjects, 147 breast cancer cases and 253 controls, were interviewed and information on lifestyles including smoking and alcohol drinking habits, menstrual and reproductive history, family history of cancer, and physical activity, was collected using a questionnaire. Erythrocyte fatty acids were measured by gas-liquid chromatography. According to the results of questionnaire survey, there was no inverse association between the frequency of fish intake and the risk of breast cancer. In pre-menopausal women, there was no significant association between DHA / AA and breast cancer risk (P for trend = 0.953). For post-menopausal women, although the first quartile of DHA / AA showed the lowest breast cancer risk, the risk was the highest in the second quartile and decreased after that. There was no association between EPA / AA and breast cancer risk in both pre- and post-menopausal women. These results suggested no inverse linear association between DHA(EPA) / AA in erythrocytes and breast cancer risk.
Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-09-06.
Abstract P4-10-01: High dose omega-3 fatty acid supplementation modulates breast tissue biomarkers in post-menopausal women at high risk for development of breast cancer
Authors: Fabian CJ , Kimler BF , Petroff BK , Zalles CM , Metheny T , Nydegger JL , Box JA , Phillips TL , Hidaka BHH , Carlson SE , deGraffenried LA , Hursting SD 
Published: August 2014

Abstract
Background: We conducted a pilot study of high dose omega-3 fatty acid (FA) supplementation in post-menopausal women to determine if risk biomarkers for breast cancer in benign breast tissue sampled by random peri-areolar aspiration (RPFNA) could be favorably modulated and to acquire preliminary data on possible mechanism of action.
Methods: 35 post-menopausal women at increased risk for breast cancer were accrued to a trial of 6-month intervention with 4 g daily of omega-3-acid ethyl esters [1.86 g eicosapentaenoic acid (EPA), 1.5 g docosahexaenoic acid (DHA)]. Subjects had RPFNA performed pre- and post-intervention and specimens evaluated for cytomorphology and proliferation (Ki-67). FA composition was determined in plasma, red blood cells, and RPFNA specimens. Additional specimens were frozen for assessment of hormones, a panel of 11 adipokines and cytokines by Luminex, and gene expression.
Results: 34 subjects completed study with specimens evaluable for change in biomarkers. The ratio of (EPA+DHA):Arachidonic Acid (AA) levels in erythrocyte phospholipid increased significantly by a median of 2.7-fold. Although there was a significant decrease in blood EPA+DHA between discontinuation at 6 months and 2 weeks later when RPFNA was performed, all ratios were above the baseline value (median 1.6-fold). There was favorable but not statistically significant modulation for cytologic evidence of atypia (53% at baseline to 41% at off-study). However, favorable modulation was exhibited for Masood score (medians of 15 to 14; p = 0.014), number of epithelial cells recovered (p = 0.019) and Ki-67 expression (medians of 1.7% to 0.75%, p = 0.036, despite 8 subjects having no Ki-67 expression at baseline). Luminex assay of serum indicated a statistically significant increase (p = 0.003) for adiponectin and decrease (p = 0.016) for TNF-alpha between baseline and off-study. For RPFNA specimens, there was a significant decrease (P = 0.001) in MCP-1 levels adjusted for protein content. By ELISA, serum high molecular weight adiponection increased (p = 0.046) and molar ratio of IGF-1:IGFBP3 decreased (p = 0.006). Note that all analyses were exploratory and without correction for multiple analyses.
Conclusion: Favorable modulation of a variety of blood and tissue risk biomarkers, including cytomorphology and proliferation, along with good tolerability suggests that high dose omega-3 FA esters should be tested further in a placebo-controlled trial.
Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-10-01.
Abstract P4-11-02: Mammary tumor formation induced by N-methyl-N-nitrosourea (MNU) is accelerated by natural and synthetic progesterone but suppressed by an anti-progesterone CDB4124
Authors: Lee O , Sullivan M , Ivancic D , Shidfar A , Wiehle R , Khan SA 
Published: August 2014

Abstract
Background: CDB4124, anti-progesterone suppresses the development of carcinogen-induced ER+/PR+ mammary tumors in rats, and may have implications for prevention and treatment of human breast cancer. We hypothesize that progesterone (P4) and medroxyprogesterone acetate (MPA) will accelerate mammary carcinogenesis induced by MNU, however CDB4124 will efficiently suppress tumor formation stimulated by progesterone.
Methods: ovary intact female Sprague Dawley rats received a single intraperitoneal injection of 50mg/kg MNU at 4-5 weeks of age. 30mg of CDB4124 and 25mg of P4 or MPA (90 release pellets, Innovative research of America, Inc) were implanted in dorsal area at 3 weeks and 4 weeks after MNU injection, respectively. 10-11 rats were used for each treatment group. Tumor incidence, latency, multiplicity, and burden were recorded weekly. 9 weeks after MNU injection all the mice were euthanized and mammary tumors and glands were fixed in 10% (v/v) neutral buffered formalin. Plasma concentrations of CDB4124 and its metabolite CDB4453 were determined by LC-MS/MS.
Results: The first tumor appeared in the control group at 5 weeks, and in the P4 and MPA treated groups at 6 weeks after MNU injection. 7 weeks after MNU injection, mammary tumor incidence of MPA and P4 treated groups were 80% and 50%, respectively compared to 30% in the control group. 9 weeks after MNU injection all MPA treated mice, 80% of P4 treated mice, and 60% of control mice developed tumors. Tumor incidence, latency, multiplicity, and tumor weight were summarized as mean ± SD in Table 1.
[Table 1] Tumor latency, incidence, multiplicity, and burden in mammary tumorsTreatmentsLatency (days)Incidence(%)MultiplicityBurden (g)Control53.7 ± 12.9602.34.25 ± 7.02P453.6 ± 9.6802.64.11 ± 5.32MPA50.8 ± 7.71002.56.02 ± 4.85P4 + CDB412459.3 ± 11.53621.38 ± 0.61MPA + CDB412454.3 ± 10.7732.84.19 ± 4.39
Tumor latency of CDB4124 treated groups was increased; tumor incidence and burden (g) of CDB4124 treated groups were decreased compared to P4 and MPA treated groups. In particular, tumor incidence and burden of CDB4124 + P4 treated group were significantly lower than those of the control group. Plasma CDB4124 and CDB4453 were 11.6 ±5.88 ng/mL and 3.4±1.68 ng/mL, respectively. Histopathology of tumors and mammary glands and immuno-histochemical evaluations of Ki67, activated caspase-3, CD34, ER, and PR are currently underway.
Conclusions: Our results indicated that natural progesterone promotes MNU- induced mammary tumor formation similar to synthetic progesterone, MPA in rats. Under this tumor permissive environment, CDB4124 provided excellent prevention efficacy, suggesting good potential as breast cancer prevention agent.
Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-11-02.
Abstract P4-12-06: Small HER2-positive breast carcinomas: Prognostic factors to consider in deciding on adjuvant trastuzumab treatment
Authors: Campiglio M , Sandri M , Sasso M , Bianchi F , Turdo F , Ménard S , Tagliabue E 
Published: August 2014

Abstract
Criteria for the decision to treat small tumors in adjuvant setting remain debatable, especially for HER2-positive tumors, since HER2 itself is associated with poor prognosis. Analysis of a large database including 503 T1 HER2-positive breast carcinomas treated with chemotherapy plus trastuzumab in Italian oncology clinics from 2006 to 2009 and, with 32 months of median follow-up and with 35 (7.0%) relapses, revealed no statistically significant differences in relapse rate among the T1a/T1micro, T1b and T1c groups, suggesting that tumor size is not associated with relapse probability in T1 HER2-positive tumors; the only clinico-pathological factors significantly associated with disease-free survival were nodal involvement (unadjusted HR = 3.4, 95%CI = 1.6-7.3, p = 0.002), loss of hormone receptor expression (HR = 3.1, 95%CI = 1.5-6.2, p = 0.002) and high tumor grade (HR = 2.9, 95%CI = 1.2-6.9, p = 0.020). Multivariate analysis indicated that patients who were both ER-negative and lymph node-positive presented the highest risk of relapse (HR = 5.5, 95%CI = 2.8-10.9, p = &lt;0.001). These results suggest that the clinical decision for adjuvant treatment of small HER2-positive tumors should be based on nodal and hormone status, not on tumor size information. While the most recent National Comprehensive Cancer Network guidelines (version 3.2012) for systemic adjuvant treatment of small HER2-positive breast cancer only suggest considering adjuvant chemotherapy treatment including trastuzumab for T1a cases with axillary node micrometastasis and for all T1b, but require such treatment for all tumors &gt;1 cm, our data identify T1 tumor ER-negativity/axillary lymph node-positivity as the crucial criteria for adjuvant treatment, independent of tumor size categories.
Supported by AIRC.
Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-12-06.