bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract LB-435: Double-blind randomized clinical trial of vitamin D3 showing effects on tissue calcitriol levels, gene expression and proliferation immunohistochemistry in prostate cancer
Authors: Wagner Dennis , Nonn Larisa , Antonio Angeline , Vaishnav Avani , Klotz Laurence , Fleshner Neil , Finelli Antonio , Trudel Dominique , van der Kwast Theodorus , Vieth Reinhold
Published: August 2012

Background: Preclinical data suggest that vitamin D plays a favourable role in prostate cancer prevention and prognosis, but its putative anticancer effects have not been validated in clinical prostate tissue samples. Here, we report the results from a randomized clinical trial (RCT). Objectives: We conducted an RCT to determine whether simple cholecalciferol (vitamin D3) consumed orally elicits effects on prostate cancer pathology in prostate tissue obtained at surgery. Methods: Prostate cancer patients (n=66) with Gleason scores 6 or 7 and scheduled for radical prostatectomy were randomized to daily vitamin D doses of 400, 10,000, or 40,000 IU for 3-8 wk before surgery. Results: Laser-capture microdissected paraffin-embedded prostate tissue showed differential expression, between benign and cancerous regions, for the cancer-related mRNAs and miRNAs, as well as for the Ki-67 proliferation marker (p0.07 among groups). The group consuming 40,000 IU/d vitamin D exhibited the highest levels of both calcitriol and its precursor as measured in frozen prostate tissue and in serum (p<0.02 each). Prostate tissue calcitriol correlated strongly with the expression of androgen receptor mRNA (r = -0.58, p=0.001), NKX3.1 mRNA (r = -0.39, p=0.02), miR-100 (r = 0.44, p=0.01), miR-125b (r = 0.37, p=0.03), Let-7a (r = 0.47, p=0.03), miR-141 (r = 0.51, p=0.002), miR-106b (r = 0.45, p=0.01), and miR-331-3p (r = 0.47, p=0.005). Automated digital analysis of immunohistochemically stained prostate cancer regions showed that prostate tissue calcitriol related negatively with Ki67 intensity (r = -0.38, p =0.02) and percent of nuclei stained strongly positive (3+) for Ki67 (r = -0.41, p=0.008). A threshold effect was evident such that patients at the highest quartile of prostate tissue calcitriol (≥ 37 pmol/kg) had significantly lower AR mRNA (p<0.001), NKX3.1 mRNA (p=0.01), and Ki67 measures (p<0.05), as well as higher levels of the correlated miRNAs (p<0.05), compared to those with lower levels. Conclusions: We provide level-1 clinical evidence that higher oral vitamin D doses produce higher levels of calcitriol within prostate tissue, and that higher prostate calcitriol relates to lower androgen receptor mRNA, higher miRNA signalling, and diminished expression of the proliferation marker Ki67 in prostate cancer. These RCT results justify a need to explore strategies that may maximize calcitriol within prostate for cancer chemoprevention.

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 LB-435. doi:1538-7445.AM2012-LB-435
Abstract LB-437: Co-amplification of ALK and c-MET in a subset of pulmonary sarcomatoid carcinoma (PSC)
Authors: Gasparini Patrizia , Sozzi Gabriella , Cavazza Alberto , Rossi Giulio , Papotti Mauro , Pastorino Ugo , Scanagatta Paolo , Nakatani Yukio , Pelosi Giuseppe
Published: August 2012

The prevalence of ALK gene alterations for targeted therapy with specific inhibitors is a largely unexplored issue in pulmonary sarcomatoid carcinoma (PSC), a life-threatening tumor subset whose treatment still is disappointing. ALK gene status by fluorescence in situ hybridization (FISH) and protein expression by immunohistochemistry (IHC) were carried out on formalin-fixed, paraffin-embedded material from 34 PSC. Fifty-one consecutive metastatic lung adenocarcinomas (MELAD) were also assessed in parallel for ALK with both FISH and IHC. While no rearrangements of ALK gene were detected in PSC, relevant amplification was identified in 6/34 (18%) tumors, while other genes known to behave as driver mutations according to an oncogene addiction criterion, such as EGFR, KRAS, HER2, BRAF, PIK3CA and CTNNB1, were consistently lacking. The percentage of amplified tumor cells ranged from 11% to 43%, with a mean gene copy gain (GCG) ± standard deviation (SD) of 7.5 ± 1.5. We have also investigated 51 MELAD observing that, while 10 of them showed the expected ALK rearrangement (p=0.012), only 1/51 showed the gene amplification (p=0.015). No cases of ALK-amplified tumors, either PSC or adenocarcinoma, expressed the relevant protein, whereas it was otherwise detectable in all ALK-rearranged adenocarcinomas. Interestingly all ALK-amplified tumors, either PSC or adenocarcinoma, showed also c-MET amplification consisting of variably sized clusters of specific signals, whereas negative tumors, regardless of histology, and ALK-rearranged adenocarcinomas did not show c-MET alteration. ALK and c-MET gene amplification clustered into a significant subset of elderly, smoking, predominantly male PSC patients, revealing significant differences with lung adenocarcinoma These encouraging data, however, need to be reinforced on a wider series of tumors in order to elucidate this issue, which could potentially lead to a truly effective treatment of these so life-threatening 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 LB-437. doi:1538-7445.AM2012-LB-437
Abstract LB-441: Mutation in SLX4 gen in a multiple case breast cancer family
Authors: de Garibay Ponce Gorka Ruiz , Díaz María Avellaneda , Gaviña Belén , Perez-Segura Pedro , Díaz-Rubio Eduardo , Caldés Trinidad , de la Hoya Miguel
Published: August 2012

Fanconi Anemia (FA) is a genetically heterogeneous, rare autosomal recessive disorder characterized by development abnormalities, bone marrow failure, and childhood cancers. Over the past few years, a common genetic basis for FA and breast and/or ovarian cancer susceptibility has been uncovered. In particular, BRCA2 (FANCD1), BRIP1 (also known as BACH1 and FANCJ), PALB2 (FANCN), and RAD51C germ-line mutations have been found to lead both to FA (when biallelic) and to increased breast and/or ovarian cancer risk (when monoallelic). Recently, two studies demonstrated that biallelic germ-line mutations in SLX4, which encodes an endonuclease, cause a previously unknown FA subtype (FA-P). Therefore, SLX4 should be considered a bona fide candidate gene for breast cancer susceptibility, as suggested in both studies. We have screened 347 non-BRCA1/BRCA2 index cases from multiple case breast and/or ovarian cancer families for point mutations in the SLX4 gene. The whole coding sequence and flanking intronic regions were analyzed using High Resolution Melting (HRM) analysis followed by direct sequencing of abnormal melting curve samples. An SLX4 missense change in c.1114C>T (p.Arg372Trp) was detected in a family with four cases of breast cancer in two generations. This mutation segregates with disease within the family and is not present in the dbSNP (build 34). Polyphen-2 and SIFT in silico analysis tools predict a deleterious effect on protein structure and multiple alignment within orthologous proteins suggests a high degree of conservation. Previous studies in FA patients have identified two different splicing mutations (c.1163+2T>A and c.1163+3dupT) that cause in frame exon 5 skipping. The resulting protein (p.Arg317_Phe387del) lacks two repeated UBZ4 domains known to interact with other endonucleases, mismatch repair proteins, and ubiquitin. It has been suggested that UBZ4 may be involved in targeting SLX4 to sites of DNA damage by binding to ubiquitinated proteins. The missense change we have found introduces a highly hydrophobic amino acid within the same region, which could affect the folding of the UBZ4 domain or disrupt a specific protein-protein interaction. Our study supports a role for SLX4 in breast cancer susceptibility. Further studies performed in larger cohorts will be necessary to determine the prevalence of SLX4 mutations among breast cancer risk factors.

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 LB-441. doi:1538-7445.AM2012-LB-441
Abstract LB-443: The Simple Western rapidly generates quantitative profiles of MAPK and PI3K proteins in clinical specimens
Authors: Fan Alice C. , Banerjee Prajna , Felsher Dean W.
Published: August 2012

In this era of targeted therapeutics, there is a need for highly sensitive and quantitative methods to measure proteins in clinical specimens to define and predict of specific therapies for patients. However, the current gold standard in proteomic analysis, Western Immuno Blotting, has many manual steps, is insensitive and provides only semi-quantitative data. Here, we present the first report of a novel automated platform, the Simple Western(TM), a novel automated nanovolume size-based protein separation platform used to quantify proteomic profiles of clinical specimens. Since the Simple Western automates all steps of proteomic analysis including sample loading, size-based protein separation, immunoprobing, washing, detection and data analysis, we are able to make up to 96 measurements in a single experiment, in a highly quantitative manner, minimizing errors due to manual variability. We were able to perform direct proteomic comparison of oral cavity tumors paired with adjacent normal tissues, acquired from the Stanford University Tissue Bank that had been preserved at -80C in Optimum Cutting Temperature Compound (OCT). Tissues were homogenized and lysed in a commercially available Bicine/CHAPS lysis buffer, denatured and loaded in duplicate in a 384-well plate at a final concentration of 1-2 mg/ml in each well. 40 nanoliters of lysate was used for each protein measurement. We measured 10 proteins (including AKT and ERK) from the mitogen activated protein kinase (MAPK) signaling pathways, normalized to loading controls (β-actin) from 4 distinct clinical specimens and 2 additional cell line controls. It took 1 hour to prepare a run and 14 hours of unattended automated machine time to generate the analyzed data. Therefore this technology enables us to report highly quantified protein profiles to the clinical team in less than 24 hours of receiving a clinical sample. Moreover, the highly quantitative nature of the measurements allowed us to determine that AKT2 levels were 80% lower in the tumor tissue as compared to their adjacent normal samples. Furthermore, we have been able to show that levels of both ERK1 and ERK2 were independently lower by 71% and 70%, respectively, in the tumor samples as compared to their corresponding adjacent normal tissues. The ability to make rapid and quantitative measurements will be useful to measure and predict responses to targeted therapeutics. The Simple Western is an extremely efficient, quantitative and high-throughput platform that can be used to rapidly generate proteomic profiles for clinical specimens for the development of novel diagnostic and predictive biomarkers.

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 LB-443. doi:1538-7445.AM2012-LB-443
Abstract LB-444: SATB2 is a novel prognostic factor in colorectal cancer
Authors: Naicker Kirsha , Brennan Donal , Klinger Rut , Ponten Fredrik , Jirstrom Karin , O'Sullivan Jacintha , Gallagher William M. , O'Connor Darran P.
Published: August 2012

SATB2 is a nuclear matrix-associated transcription factor that orchestrates gene expression in a tissue-specific manner by regulating higher order chromatin structure. Using antibody-based screening of 48 normal human tissues and 20 cancer types, we identified SATB2 as a protein almost exclusively expressed in gastrointestinal tissue. Differential expression of SATB2 was observed in colorectal cancer, with loss of expression occurring along the adenoma-carcinoma sequence. We further confirmed this observation at the mRNA level in 10,533 colorectal cancer samples. Additionally, SATB2 expression was markedly decreased in a metastatic variant of SW480 colon cancer cells and siRNA knockdown of SATB2 expression in parental SW480 cells increased their growth and migratory capacity. Ectopic expression of SATB2 in the metastatic variant reversed the observed phenotype. Using tissue microarray and automated image analysis of SATB2 expression in colorectal cancers (n=309), SATB2 was demonstrated by multivariate Cox regression analysis to be an independent predictor of disease-specific survival (HR=0.52, 95% CI 0.32-0.83, p=0.006) and loss of SATB2 expression significantly correlated with poor differentiation (p=0.001). SATB2 mRNA levels were examined in a second cohort (n=290) and again, SATB2 was demonstrated to be an independent predictor of disease-specific survival (HR=0.40, 95% CI 0.18-0.92, p=0.031). Interestingly, in colorectal cancer patients, SATB2 levels significantly correlate with CD3+ T-cell infiltrates in the tumours (p=0.006) and inversely correlate with COX2 expression (p=0.019). Gene set enrichment analysis in two independent colorectal cancer cohorts (n>500) revealed that SATB2 low tumours demonstrate altered immune signalling with significant increases in IFN-gamma(p=0.001), IL6 (p=0.021) TFGβ (p<0.001). Furthermore, in patients with inflammatory bowel disease (n>1000), we observed a significant correlation between loss of SATB2 expression and occurrence of future cancers (p=0.013). We therefore postulate that SATB2 acts as a master regulator of the inflammatory response in the gut and loss of expression is significantly associated with the progression of colorectal 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 LB-444. doi:1538-7445.AM2012-LB-444
Abstract LB-446: Target-based therapeutic matching in patients with PIK3CA mutations and PTEN aberrations
Authors: Janku Filip , Wheler Jennifer J. , Piha-Paul Sarina A. , Stepanek Vanda M. , Fu Siqing , Hong David S. , Naing Aung , Falchook Gerald S. , Moulder Stacy L. , Luthra Rajyalakshmi , Tsimberidou Apostolia M. , Kurzrock Razelle
Published: August 2012

Background: Activating mutations of PIK3CA or loss of PTEN function result in increased PI3K/AKT/mTOR signaling and may predict sensitivity to drugs targeting the PI3K/AKT/mTOR pathway, while simultaneous KRAS mutations may mediate resistance. Methods: From 10/2008, available tumor samples from advanced cancer patients referred to the Phase I Clinical Trials Program were screened for PIK3CA mutations and PTEN aberrations (loss of expression on immunohistochemistry or mutation) in a CLIA-certified laboratory. Patients with tumors carrying PIK3CA mutations and/or PTEN aberrations were treated, whenever possible, with agents targeting the PI3K/AKT/mTOR pathway. Results: Overall, 1,540 patients were tested, and 285 (19%) had PIK3CA mutations (n=141), PTEN aberrations (n=130), or a simultaneous PIK3CA mutation and PTEN aberration (n=14). Of these 285 patients, 131 (46%) were treated in clinical trials containing a PI3K/AKT/mTOR inhibitor (median age, 56; median number of prior therapies, 3). Of these 131 patients, 26 (20%, 95% CI 0.14-0.27) achieved a partial response (PR) (4/21 breast cancers; 4/19 endometrial cancers; 3/13 ovarian cancers; 4/9 head and neck cancers; 4/10 cervical cancers; 2/9 renal cancers; 2/4 salivary gland cancers; 1/4 sarcomas; 1/3 non-small cell lung cancers; 1/2 melanomas). Patients without known PIK3CA mutations and/or PTEN aberrations treated on the same protocols had a PR rate of 6% (26/458; 95% CI 0.04-0.08, p=0.0001). Treated patients with wild-type KRAS had a higher PR rate of 20% compared to 8% in patients with simultaneous KRAS mutations (p=0.05). Conclusion: Heavily pretreated patients with PIK3CA mutations and/or PTEN aberrations had higher PR rates on protocols with PI3K/AKT/mTOR inhibitors than patients without these aberrations (20% vs 6%; p = 0.0001). Simultaneous KRAS mutations were associated with a lower PR rate. Screening for PIK3CA mutations and PTEN aberrations merits further evaluation in making treatment decisions.

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 LB-446. doi:1538-7445.AM2012-LB-446
Abstract LB-457: Multiplexing protein and DNA biomarkers on a single analytical platform
Authors: Li Shuqiang , Campo Maria F. , Fernandez Cecilia A. , Shuber Anthony P.
Published: August 2012

We have recently developed a non-invasive assay that combines protein and DNA biomarkers for triaging bladder cancer patients and demonstrated that this multi-analyte approach improves clinical performance. This assay combines detection in urine of Matrix Metalloproteinase-2 (MMP-2) protein, hypermethylation of two DNA markers (Twist1 and Nid2) and mutation detection of the FGFR3 gene. To eliminate the need to perform separate analytical procedures for protein and DNA, and to decrease assay complexity and cost while increasing assay throughput, we are now developing a method for simultaneous detection of protein and DNA biomarkers on a qPCR platform. To this end, we have identified and characterized six DNA aptamers that specifically bind Matrix Metalloprteinase-2 (MMP-2). DNA aptamers are single stranded DNA oligos that can bind specifically to their target molecules, including proteins. Once bound to the protein, these aptamers can then be used as templates for quantitative PCR-mediated protein detection. We have developed qPCR assays for each of the six MMP-2-specific aptamers, and demonstrate detection limits for each aptamer as low as 5x102 copies. Kinetic and equilibrium analysis was used to determine the KD constant for the aptamers. High affinity interactions were observed for all aptamers, with two aptamers having the highest affinity and calculated KD values of 15 and 10nM. Using one of these aptamers, we bound MMP-2 protein to the aptamer in solution and then immunoprecipitated the protein/aptamer complexes using biotin-labeled MMP-2-specific antibodies. Immunoprecipitated complexes were then quantitated using the aptamer-specific qPCR assay. In this study, we show that by qPCR, we can efficiently detect 10-12 mol/L of MMP-2 using the aptamer as the detection molecule. This level of MMP-2 protein detection is equivalent to commercially available ELISA assays. To further improve performance and to take advantage of the known specificity of aptamers, we intend to develop a system that uses one aptamer for detection and a second biotin-labeled aptamer for immunoprecipitation. We subsequently demonstrate the ability to multiplex the aptamer qPCR assay with our qPCR assay for FGFR3 and intend to add the other DNA markers into this multiplex system in the future. This is the first demonstration of simultaneous detection of protein and DNA biomarkers using a single analytical platform.

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 LB-457. doi:1538-7445.AM2012-LB-457
Abstract LB-462: AGO2 PAR-CLIP suggests a shift in specific miRNA target spectra upon modulation of EWS-FLI1 in Ewing's sarcoma
Authors: Herrero-Martin David , Kauer Max , Sienski Grzegorz , Fourtouna Argyro , Kralovics Robert , Kovar Heinrich
Published: August 2012

The chimeric oncoprotein EWS-FLI1 is the driving pathogenic force in Ewing's sarcoma family tumors (ESFT). As an aberrant ETS transcription factor it directly deregulates a multitude of genes related to proliferation control and differentiation. Recently, we reported on the EWS-FLI1 microRNA (miRNA) signature based on knockdown experiments in ESFT cell lines and on differential expression in primary tumors versus mesenchymal stem cells. In order to define the mRNA targets of EWS-FLI1 regulated miRNAs, we now performed pull-down experiments of AGO2 complexes after photoactivated ribonucleoside-enhanced cross-linking followed by NGS sequencing of associated mRNAs (PAR-CLIP). We obtained 92 Mio high quality reads of which 63 Mio were aligned to the reference genome. The genomic localization of these sequences demonstrated the specific enrichment of 3'UTRs (>17 fold). Among obtained reads, T to C conversions were highly enriched consistent with crosslinking via 4-thiouridine to AGO2 containing complexes. PAR-CLIP results indicate AGO2 binding next to miRNA seed sequences for 4695 and 5160 genes associated with 1876 and 1860 miRNAs in the presence and absence of EWS-FLI1, respectively. Strikingly, for individual miRNAs the mRNA target spectra drastically changed upon EWS-FLI1 knockdown. For instance, for the two top EWS-FLI1 signature miRNAs hsa-mir-145 and hsa-mir 424 a shift from genes involved in cell death and differentiation to protein activator activity, and from protein degradation and cell cycle control to protein kinases and protein phosphorylation was observed, respectively. Of note, a target shift was also observed for miRNAs that did not change in response to EWS-FLI1 knockdown. In addition for many specifc targets no correlation was observed between mRNA abundancy and the level of association with AGO2 as reflected by read counts. These data suggest that EWS-FLI1 perturbs post-translational gene regulation by changing access of the RISC complex to its target mRNAs. Supported by EC-FP7 grant 259348 “ASSET”.

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 LB-462. doi:1538-7445.AM2012-LB-462
Abstract LB-467: miR-135b and miR-210 induce invasion and distant metastasis by targeting SIAH1 and SETD2 in colorectal cancer
Authors: Mudduluru Giridhar , Lunavat Taral R. , Scharp Maike , Muppala Santoshi , Patil Nitin , Leupold Jörg H. , Abba Mohammed , Allgayer Heike
Published: August 2012

The metastatic spread of tumor cells is ultimately responsible for the vast majority of cancer related mortality. In the attainment of a metastatic phenotype, several intricately interwoven processes are involved that lead to uncontrolled proliferation, loss of intracellular adhesions, extracellular matrix degradation, intravasation and dissemination into distant organs. Therefore, an improved understanding of molecular mechanisms which regulate metastatic transformation and progression of the cancer cells is of paramount importance. In this present study we performed miRNA-microarray analysis of six patient samples comprising normal, primary tumor and distant metastasis tissues, where we found miR-135b, miR-210, miR-378, miR-19a to be significantly up-regulated in both tumor and metastasized tissues as compared to the respective normal equivalents. We validated miR-135b and -210 in a series of 30 colorectal patients, in which we were able to demonstrate an increased expression in tumor as compared to normal tissues (p=0.008 and p=0.03, respectively). Forced in vitro expression of these miRNAs resulted in increased migration and invasion of Rko, SW480 and SW620 colorectal cancer (CRC) cell lines. Additionally, these cells manifested a gain of mesenchymal marker (N-cadherin) and a loss of epithelial marker (E-cadherin) expression at the protein level. Furthermore, of the several predicted targets identified, through luciferase, RT-PCR and Western blot analysis we found SIAH1 and SETD2 to be regulated by miR-135b and -210, respectively. Furthermore, in vivo, chorioallantoic membrane (CAM) metastasis assays revealed that miR-135b and miR-210 significantly induce distant metastasis to the lungs and liver (p=0.05). Our preliminary findings suggest that miR-135b and 210 play important roles in inducing tumor progression and may be useful as potential targets for mitigating metastatic spread

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 LB-467. doi:1538-7445.AM2012-LB-467
Abstract LB-469: An Epstein-Barr virus-encoded microRNA miR-BART5 upregulates human cellular microRNA miR-146a and downregulates IRAK1/NFκB in stomach cancer cells
Authors: Kim Dong Ha , Rho jin Kyung , Woo Jun Hee , Lee Byung Lan , Kim Chul Woo , Kim Woo Ho , Chang Mee Soo
Published: August 2012

Epstein-Barr virus (EBV) contributes to the development of the specific subsets of stomach cancer, nasopharyngeal cancer and malignant lymphoma etc. MicroRNAs (miRNAs) have been implicated in the regulation of various pathobiological processes in cancer. Recently, EBV-encoded viral miRNAs have been disclosed, whereas the functions of which remain largely unknown. A notable example of this is the human miRNA, which is targeted by viral miRNA. In this study, we investigated EBV-encoded viral miRNA miR-BART5/human cellular target interaction. The EBV miR-BART5 in EBV-infected stomach cancer cell line SNU719 was highly expressed comparing with that in EBV-infected lymphoma cell lines, Raji and Jijoye. In addition, EBV-infected stomach cancer cell revealed high level of human cellular miR-146a comparing with EBV-negative stomach cancer cells SNU601 and SNU216. Either miR-BART5 inhibitor product (anti-miR-BART5) or miR-BART5 precursor product (pre-miR-BART5) was transfected into EBV-infected stomach cancer cell, or EBV-negative stomach cancer cell, respectively. As a result, the expression of human cellular miR-146a was decreased in EBV-infected stomach cancer cell transfected with anti-miR-BART5 comparing with that in its original cell, while miR-146a was increased in EBV-negative stomach cancer cell transfected with pre-miR-BART5. As a predicted target of miR-146a, based on predicted base pairing using miRBase online program, we focused on interleukin-1 receptor-associated kinase 1 (IRAK1). Once EBV-negative stomach cancer cell was transfected with pre-miR-BART5, IRAK1 protein level was reduced by 50%, and nuclear factor-kappaB (NF-κB) (p65) was decreased. To check miR-BART5 influences on cellular biologic properties, when pre-miR-BARF5 was transfected into EBV-negative stomach cancer cell, invasion capacity of cancer cell was suppressed with a statistical significance. In conclusion, miR-BART5 could inhibit invasion of stomach cancer cell, which is in part due to the upregulation of miR-146a and downregulation of IRAK-1/ NF-κB signal pathway, offering clinicopathologic entity with a better prognosis to EBV-infected stomach 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 LB-469. doi:1538-7445.AM2012-LB-469
Abstract LB-47: The Prostate Net: We Inform To Fight
Authors: Arias Janet
Published: August 2012

The Prostate Net (TPN) is a non-profit patient education and advocacy organization dedicated to serving Prostate Cancer patients and their caregivers. Using the experience gained as a marketing professional and as a 16-year Prostate cancer survivor, TPN founder and president Virgil Simons has built an international organization that uses websites, 800 numbers, email and personal team counselors, newsletters and public forums to address disease risk awareness and early intervention. TPN's core objectives are to (1) Educate consumers most at-risk for a diagnosis of prostate cancer; (2) Inform the community on co-morbid diseases and conditions of negative impact; (3) Motivate consumers to make informed choices about healthcare and lifestyle management; (4) Lay the foundation for on-going healthcare information dissemination and interaction between the community and medical centers; and (5) Create an interactive network to maximize broad scale, mass communications of actionable health messages.

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 LB-47. doi:1538-7445.AM2012-LB-47
Abstract LB-473: The HDAC inhibitor entinostat targets erbB2/erbB3 receptors via functional cooperation of miR-125a, miR-125b, and miR-205 in breast cancer cells
Authors: Wang Shuiliang , Huang Jingcao , Lee Choon-Kee , Liu Bolin
Published: August 2012

Breast cancer is highly heterogeneous and has been classified into five distinct subtypes, among which erbB2-overexpressing (erbB2+) and basal breast cancers have the least favorable prognosis and express erbB receptors that are excellent targets for anti-cancer therapies. We previously reported that the class I HDAC inhibitor, entinostat (also known as MS-275, SNDX-275, Syndax Pharmaceuticals, Inc., Waltham, MA) selectively inhibited proliferation and induced apoptosis in erbB2+ breast cancer cells. Entinostat preferably targeted erbB2 and erbB3 receptors rather than the epidermal growth factor receptor (EGFR). Here, we study the molecular mechanism through which entinostat downregulates erbB2/erbB3 and promotes the breast cancer cells undergoing apoptosis. We showed that entinostat dramatically reduced the protein levels of erbB2/erbB3, whereas it had no significant effect on their mRNA expression in MDA-MB-453 and BT474 breast cancer cells. In addition, entinostat specifically decreased the endogenous, but not the exogenous erbB2/erbB3, suggesting that it did not promote erbB2/erbB3 protein degradation. Therefore, we hypothesizes that entinostat reduces erbB2/erbB3 receptors through transcription-independent mechanism; it may inhibit erbB2/erbB3 protein translation via induction of specific miRNA. Quantitative real-time (qRT) PCR revealed that entinostat significantly upregulated the expression of three erbB2/erbB3-targeting miRNAs, miR-125a, miR-125b, and miR-205 in erbB2+ breast cancer cells. Specific miRNA inhibitors were used to determine whether these miRNAs played a causal role in entinostat-induced downregulation of erbB2/erbB3 and apoptosis. While inhibition of any one of the three miRNAs failed to block entinostat action, simultaneous inhibition of any two of them significantly abrogated entinostat-induced erbB2/erbB3 reduction and apoptosis. Collectively, these data demonstrate that entinostat targets erbB2/erbB3 receptors via functional cooperation of miR-125a, miR-125b, and miR-205. Our findings suggest that miRNA-mediated epigenetic regulation may represent a novel strategy targeting erbB2/erbB3 to treat the aggressive erbB2+ breast cancer. The combinations of entinostat and erbB2-targeted therapy may be more efficacious for breast cancer patients whose tumors overexpress erbB2. This hypothesis will be directly tested by a recently initiated clinical study determining the activity of entinostat in combination with lapatinib in breast cancer patients that are erbB2+ and progressed on trastuzumab.

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 LB-473. doi:1538-7445.AM2012-LB-473
Abstract LB-48: The survivorship and advocacy task force model at Breast Cancer Connections
Authors: Brain Susie
Published: August 2012

This poster demonstrates that an advocacy program can facilitate effective collaboration between a non-profit cancer support organization and the cancer research community. The evolution and scaling of the Survivorship and Advocacy Task Force Model is illustrated so that it can be replicated by other non-profit disease support organizations. Breast Cancer Connections (BCC), is a non-profit breast cancer resource center serving people touched by breast cancer in the San Francisco Bay Area and globally via the Internet and the telephone Helpline. BCC provides personalized information, support, and connections to other local and national resources in an atmosphere of warmth, sensitivity, and understanding. BCC was founded in 1993, by a surgeon and her patient who saw a need to augment what was being offered through the medical model. With more than 35 programs and services, all offered at no charge, BCC remains a vital and unique resource. Prompted by the requirement for patient advocate and survivor involvement in the research grant application process (California Breast Cancer Research Program, DoD, Susan G. Komen for the Cure) as well as by the increasing needs of breast cancer survivors, BCC formed the Survivorship and Advocacy Task Force (SATF) in 2008. The dual purpose of the SATF is to provide input and review to researchers applying for research grants and to assist BCC in developing services and programs that will improve the lives of breast cancer survivors. Task Force members who participate in reviewing research grants must be active research advocates and have formal training in the language and concepts of breast cancer research (such as Project LEAD Institute) or a Ph.D. in a discipline relevant to breast cancer research.The poster author chairs the BCC's SATF with five active members. The Task Force's efforts include meeting the PI to hear about the research, providing input to the PI on the research from the patient/survivor perspective, reviewing the grant proposal, refining the language of the lay abstract, writing a letter of support to accompany the grant proposal, and participating in the research if the research application is accepted. Current collaborators include breast cancer researchers from Stanford University, Palo Alto Medical Foundation, and University of California, San Francisco. As funding agencies increase their requirements for advocate involvement and as awareness of the importance of survivorship continues to grow, requests for SATF participation continues to increase.

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 LB-48. doi:1538-7445.AM2012-LB-48
Abstract LB-484: Molecular regulation of HIF-1α by CDK1 and the cell cycle
Authors: Warfel Noel A. , El-Deiry Wafik S.
Published: August 2012

The hypoxia-inducible factor 1 (HIF-1) transcription factor is a critical mediator of the cellular response to oxygen deprivation. HIF-1α is frequently overexpressed in human cancers due to intratumoral hypoxia as well as genetic alterations that promote HIF-1α synthesis or inhibit its degradation (i.e. gain of function mutations in oncogenic pathways, such as RAS and PI3K, or loss of function mutations to the tumor suppressor, VHL, respectively). Under hypoxic conditions, HIF-1α is stabilized and translocates to the nucleus where it activates the transcription of genes that promote angiogenesis, cell proliferation/survival, and metastasis, among other processes. While the role of HIF-1 in the regulation of the cell cycle has been well studied, the effect of the cell cycle stage on HIF-1 expression and activity remains unknown. To address this question, HCT116 colon cancer cells were synchronized in G1, S, or G2/M-phase and exposed them to hypoxia (0.2% O2). Interestingly, HIF-1α expression was nearly abolished upon G2/M-phase arrest, and a significant increase was observed following arrest in S-phase. Therefore, cell cycle position influences the expression of HIF-1α, independent of oxygen concentration. Previous data in our lab indicates that knockdown of CDK1 decreased HIF-1α expression under hypoxic conditions. To further characterize this effect, we demonstrate that acute inhibition of CDK1 kinase activity with small molecules significantly decreases the stability of HIF-1α at the protein level, without affecting HIF-1α mRNA levels. Furthermore, cycloheximide chase experiments reveled that HIF-1α protein levels decreased at a significantly faster rate in the presence of a CDK1 inhibitor, R0-3306, (t1/2 = 38.2 ± 6.1 min) compared to controls (t1/2 = 54.6 ± 3.2 min). These results indicate that the half-life of HIF-1α is significantly decreased upon inhibition of CDK1. Taken together, these data indicate that CDK1 activity regulates the stability of HIF-1α, and suggests that CDK1 activity stabilizes HIF-1α through a post-translational modification that promotes its stability. Thus, we have discovered that CDK1 and the cell cycle position regulate the expression of HIF-1α. There is a subset of tumors that express HIF-1α under normoxic conditions, driving tumor progression and therapeutic resistance. Therefore the identification of alternative mechanisms that regulate HIF-1α expression would be extremely valuable in the clinical setting, as well as to aid the basic understanding of the role of HIF-1 in tumorigenesis.

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 LB-484. doi:1538-7445.AM2012-LB-484
Abstract LB-485: Estrogen receptor alpha activation is mediated by both the MAPK and PI3K phosphorylation pathway in hypoxia
Authors: Park Joon Woo , Lee Youngjoo
Published: August 2012

The estrogen receptor (ER) plays an important role in breast cancer development and progression. Hypoxia was shown to modulate the level of ERa expression and induce ligand-independent transcriptional activation of ERa, which may be intimately associated with the biology of breast carcinomas. Given that phosphorylation affects the transcriptional activity and stabilization of ERa, we examined the changes in phosphorylation of ERa under hypoxic conditions. Hypoxia induced phosphorylation of ERa at serine residues 118 and 167 in the absence of estrogen. Phosphorylation-defective ERa mutants with serine-to-alanine replacements at residues 118 and 167 had impaired hypoxia-induced ERa activation, showing that serine residues 118 and 167 are involved in hypoxia-induced ERa activation. The hypoxia-induced ERa-mediated transcriptional response was dependent on both the ERK1/2 MAPK and PI3K pathways, but not the p38 pathway, as assessed using chemical inhibitors. These data show that ERa phosphorylation via both the MAPK and PI3K pathways is one mechanism leading to ERa activation under hypoxia.

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 LB-485. doi:1538-7445.AM2012-LB-485
Abstract LB-487: Pim protein kinase inhibitors induce the unfolded protein response and sensitize prostate cancer cells to killing by ABT-737-mediated apoptosis
Authors: Song Jin H. , Kraft Andrew S.
Published: August 2012

Pim serine/threonine kinases contribute to prostate tumorigenesis and therapeutic resistance, yet anticancer efficacy of Pim kinase inhibitors on prostate cancer is unknown. We demonstrate for the first time that a genetically engineered decrease in Pim kinase levels or the addition of Pim kinase small molecule inhibitors (e.g. SMI-4a) to human prostate cancer cells, including LNCaP and PC-3, causes ER stress and activates the unfolded protein response (UPR) stimulating increases in eIF-2α phosphorylation, ATF-4, CHOP proteins, and cleavage of XBP-1. Because (1) Bcl-2 family proteins reside on the ER lumen and play an important role in regulation of XBP-1 splicing, (2) Bcl-2 family members are over expressed in prostate cancer to enhance Pim kinase anti-cancer activity, and (3) Pim kinase inhibitors decrease Mcl-1 and increase Noxa protein, we treated prostate cancer cell lines with SMI-4a plus the Bcl-2 family antagonist ABT-737, a small molecule antagonist of Bcl-2 family members. Strikingly, the addition of ABT-737 to Pim inhibitors triggered a robust apoptosis of prostate cancer cells in vitro and in vivo. Pim inhibitors decreased levels of the Bcl-2 family member Mcl-1, both by blocking 5′-cap dependent translation and decreasing protein half life. Additionally, Pim inhibition transcriptionally increased levels of the BH3 protein Noxa by activating the UPR, lead to eIF2α phosphorylation and increased expression of CHOP. Increased levels of Noxa also inactivated the remaining levels of Mcl-1 protein activity. Notably, these specific protein changes were essential to the apoptotic process because ABT-737 did not inhibit Mcl-1 protein activity and Mcl-1 overexpression blocked the apoptotic activity of ABT-737. We find that the Pim kinase inhibitors and ABT-737 are highly synergistic in killing prostate cancer both in vitro and when used together in a subcutaneous animal model of prostate cancer therapy. Our results therefore suggest that this combination treatment could be developed as a potential therapy for human prostate cancer where overexpression of Pim kinases and antiapoptotic Bcl-2 family members drives tumor cell resistance to current anticancer therapies.

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 LB-487. doi:1538-7445.AM2012-LB-487
Abstract LB-489: MDM2 alternative splicing: fine-tuning the p53 damage response
Authors: Jacob Aishwarya G. , Singh Ravi K. , Bebee Thomas W. , Chandler Dawn S.
Published: August 2012

Cell survival is dependent upon precise control of the activity of the tumor suppressor p53. p53 is a transcription factor and is known to control a number of biological processes including cell cycle arrest and apoptosis by transactivating downstream genes. Several proteins are reported to affect p53 activity but the murine double-minute protein, MDM2, and a closely related family member, MDM4, are two of the most critical regulators of p53. In previous work, we have identified alternative splicing of both the MDM2 and MDM4 genes in response to UV damage implicating alternative splicing in the cell's normal surveillance mechanism. However, the role of the splice variants in the DNA damage pathway remains elusive. To determine the role the alternative splice forms play in the p53 damage response, overexpression of the UV-induced isoforms, MDM2-ALT1 and MDM4-ALT2, was performed. Both MDM2-ALT1 and MDM4-ALT2 are capable of negatively regulating the full-length MDM2 and MDM4 full-length proteins thereby increasing p53 levels. Interestingly, each splice variant activates a distinct and unique subset of p53 target genes. An additional type of cellular stress, hypoxia, induces alternative splicing of MDM2 as well, but generates a different splice form known as MDM2-ALT2. This splice form activates a third unique profile of p53 target genes. Hence, the splicing status of the MDM2 and MDM4 genes is capable of dictating unique p53 programs. We are also currently investigating p53-independent functions for the damage-induced splice variants to better understand their role in the damage response and tumorigenesis. This work elucidates regulated splicing as a novel mechanism by which cellular injury can control the activity of p53 within the cell. By producing specific alternatively spliced variants of both MDM2 and MDM4, yet another layer of p53 regulation is initiated in order to fine-tune the cell's response to damage. Because MDM2 and MDM4 alternative splice variants are characteristic of numerous cancer types, this work indicates that the status of MDM2 and MDM4 splicing could be a consideration in determining treatment options for cancers for which p53 activation is a therapeutic strategy.

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 LB-489. doi:1538-7445.AM2012-LB-489
Abstract LB-49: Surviving by empowerment
Authors: Togbo Judy
Published: August 2012

Men's Health Network (MHN) is a national non-profit organization whose mission is to reach men, boys, and their families where theylive, work, play, and pray with health prevention messages and tools, screening programs, educational materials, advocacy opportunities, and patient navigation. MHN was founded in 1992 by a group of men's activists, therapists, and writers interested in improving the health and well-being of men, boys and families. It is a nonprofit incorporated in the District of Columbia. MHN has also been designated as a tax exempt organization under Section 501(c)(3) of the U.S. Internal Revenue Service code. MHN has received the highest rating (4 stars) from Charity Navigator. Men's Health Network (MHN) is undertaking a series of free state symposia on the health of men and their families around the country. MHN is presenting these symposia as an effort to educate health professionals and the public-at-large about significant family health issues. The symposia create awareness of such issues with state policy leaders, elected officials, and healthcare leaders as they deal with the reality of rising health care costs. These state symposia also create opportunities for additional networking for community outreach organizations throughout the state, as well as encourage preventative health measures and physician and patient dialogue. Among those invited to participate are: local, regional, and state public health department officials, key legislators, advocacy groups, medical associations, patient advocacy organizations, churches/parishes, community health leaders, and healthcare providers.

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 LB-49. doi:1538-7445.AM2012-LB-49