bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3140: Constitutive and regulated expression of GPR109A by DNA methylation and IFN-γ in human colon carcinoma cells
Authors: Bardhan Kankana , Ganapathy Vadivel , Liu Kebin 
Published: August 2012

Abstract
Butyrate, one of the short chain fatty acids generated in the colon by microbiota from dietary fiber, has shown both preventative and therapeutic potential to counteract inflammatory disease and inflammation-mediated colorectal cancer. GPR109A, a recently identified receptor for butyrate, is highly expressed in normal colonic tissue, but is often down-regulated in human colon carcinoma cells. The molecular mechanisms underlying regulation of GPR109A expression in human colon carcinoma cells is unknown. Here we demonstrated that although there is no CpG island in the human GPR109A promoter region, the GPR109A promoter is hypermethylated in human colon carcinoma cell lines and in colon carcinoma tissue specimens. Interestingly, despite the hypermethylation of the promoter, IFN-γ, a cytokine secreted by activated cytotoxic T lymphocytes (CTL), effectively up-regulated GPR109A expression in human colon carcinoma cells, and this regulation is mediated at the transcriptional level. Furthermore, we demonstrated that the IFN-γ-activated pSTAT1 regulates GPR109A expression by directly binding to the Gamma Activation Site (GAS) elements in the GPR109A promoter region in human colon carcinoma cells. At the functional level, IFN-γ-mediated up-regulation of GPR109A resulted in increased sensitivity of the tumor cells to butyrate-induced cell death and increased level of histone acetylation in human colon carcinoma cells. In summary, our data demonstrated that although GPR109A can be constitutively silenced by DNA methylation in human colon carcinoma cells, tumor-infiltrating CTLs might use secretion of IFN-γ to up-regulate GPR109A as a mechanism to re-activate GPR109A, thereby restoring functional response of colon carcinoma cells to butyrate in the tumor microenvironment.
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 3140. doi:1538-7445.AM2012-3140
Abstract 3147: Differential epigenetic regulation of TOX subfamily high mobility group box genes in lung and breast cancers
Authors: Tessema Mathewos , Yingling Christin M. , Grimes Marcie J. , Thomas Cynthia L. , Liu Yushi , Leng Shuguang , Joste Nancy , Belinsky Steven 
Published: August 2012

Abstract
Aberrant cytosine methylation affects regulation of hundreds of genes during cancer development. In this study, a novel aberrantly hypermethylated CpG island in cancer was discovered within the TOX2 promoter. TOX2 was unmethylated in normal cells but 28% lung (n = 190) and 23% breast (n = 80) tumors were methylated. Expression of two novel TOX2 transcripts identified was significantly reduced in primary lung tumors than distant normal lung (p &lt; 0.05). These transcripts were silenced in methylated lung and breast cancer cells and 5-Aza-2-deoxycytidine treatment re-expressed both. Extension of these assays to TOX, TOX3, and TOX4 genes that share similar genomic structure and protein homology with TOX2 revealed distinct methylation profiles by smoking status, histology, and cancer type. TOX was almost exclusively methylated in breast (43%) than lung (5%) cancer, whereas TOX3 was frequently methylated in lung (58%) than breast (30%) tumors. TOX4 was unmethylated in all samples and showed the highest expression in normal lung. Compared to TOX4, expression of TOX, TOX2 and TOX3 in normal lung was 25, 44, and 88% lower, respectively, supporting the premise that reduced promoter activity confers increased susceptibility to methylation during lung carcinogenesis. Genome-wide assays revealed that siRNA-mediated TOX2 knockdown modulated multiple pathways while TOX3 inactivation targeted neuronal development and function. Although these knockdowns did not result in further phenotypic changes of lung cancer cells in vitro, the impact on tissue remodeling, inflammatory response, and cell differentiation pathways suggest a potential role for TOX2 in modulating tumor microenvironment.
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 3147. doi:1538-7445.AM2012-3147
Abstract 3151: Molecular and clinicopathological impact of miR-375 deregulation in prostate cancer
Authors: Nolte Elke , Wach Sven , Stoehr Robert , Hartmann Arndt , Szczyrba Jaroslaw , Graesser Friedrich , Wieland Wolf , Wullich Bernd , Taubert Helge 
Published: August 2012

Abstract
Background: Prostate cancer is a leading cause of tumor mortality. In order to identify the underlying mechanisms we have analyzed microRNA (miRNA) expression profiles of primary prostate cancers and non-cancer prostate tissue. As recently reported by our group, miR-375 was found to be upregulated in prostate cancer using deep sequencing and microarray analyses. We now set out to study whether there is an association between miR-375 expression and the expression of different gene members of the epithelial-mesenchymal transition (EMT) pathway. Material and Methods: Expression of miR-375 and genes of the EMT pathway (ZEB1, ZEB2, N-Cadherin) was measured by qRT-PCR in 26 pairs of prostate cancer and corresponding non-tumor prostate tissue. Bivariate correlation analyses of miR-375 and EMT members as well as for miR-375 and clinicopathological features were carried out. Results: MiR-375 showed a significant upregulation in prostate cancer using qRT-PCR (p=0.0019, paired student's t-test), whereas ZEB1, ZEB2, and N-Cadherin were consistently found downregulated (p&lt;0.05, paired student's t-test). Applying bivariate correlation analysis, a negative correlation of miR-375 expression to N-Cadherin, ZEB1, and ZEB2 could be documented (p&lt;0.05, Spearman Rho test). Furthermore, we could detect an inverse correlation of miR-375 expression to patient age (p=0.002, Spearman Rho test). Remarkably, miR-375 expression higher than median is significantly correlated to a lower tumor grade (p=0.011, Fisher's exact test). Conclusion: Expression of miR-375 is inversely correlated to the expression of N-Cadherin, ZEB1, and ZEB2. As SLUG is the only member of the EMT pathway, which is a predicted target of miR-375, we suppose SLUG to constitute the putative link between miR-375 and EMT. Furthermore, our findings show that expression of miR-375 is correlated with patient age and tumor grade.
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 3151. doi:1538-7445.AM2012-3151
Abstract 3152: Molecular networks regulated by tumor suppressive microRNA-1 and microRNA-133a in head and neck squamous cell carcinoma
Authors: Nohata Nijiro , Hanazawa Toyoyuki , Kinoshita Takashi , Kikkawa Naoko , Fuse Miki , Chiyomaru Takeshi , Yoshino Hirofumi , Enokida Hideki , Nakagawa Masayuki , Okamoto Yoshitaka , Seki Naohiko 
Published: August 2012

Abstract
(Background) Head and neck squamous cell carcinomas (HNSCC) represent 6% of all cancers. Despite of considerable advances in surgery, radiotherapy and chemotherapy, the overall 5-year survival rate for patients is less than 40% in advanced stage. The understanding of the new molecular networks of HNSCC oncogenesis by recent genome analysis would be helpful in improving diagnosis and therapy of the disease. Our expression signatures of human cancer including HNSCC revealed that the expression of microRNA-1 (miR-1) and microRNA-133a (miR-133a) were significantly reduced in cancer cells. In human genome, miR-1 and miR-133a located same chromosomal regions (miR-1-2 and miR-133a-1 on 18q11.2, and miR-1-1 and miR-133a-2 on 20q13.33) called cluster. Previously, our group reported that miR-1 and miR-133a function as tumor suppressors in several types of cancers including HNSCC. In this study, we identify the novel molecular networks regulated by miR-1 and miR-133a commonly in HNSCC. (Methods) Genome-wide gene expression analysis was performed to identify the molecular networks of miR-1 and miR-133a by microarray technique. A luciferase reporter assay was used to identify the actual binding site between miR-1 and miR-133a and these candidate target genes. Cell proliferation, migration and invasion assays were performed to investigate the functional significance of target genes in HNSCC cell lines. (Results) Genome-wide molecular targets search and luciferase reporter assay showed that transgelin-2 (TAGLN2), prothymosin-alpha (PTMA) and purine nucleoside phosphorylase (PNP) were directly regulated by miR-1 and miR-133a commonly. Silencing of these genes studies demonstrated significant inhibition of cell proliferation, migration and invasion in HNSCC cells. (Conclusions) TAGLN2, PTMA and PNP were directly regulated by tumor suppressive miR-1 and miR-133a commonly. These genes may function as oncogenes contributed to cell proliferation, migration and invasion in HNSCC. Tumor suppressive miR-1 and miR-133a and target oncogenes may provide new insights into the mechanisms in cancer. Our findings have therapeutic implications and may be exploited for future HNSCC treatments.
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 3152. doi:1538-7445.AM2012-3152
Abstract 3155: High-throughput profiling of miRNA expression in plasma and cerebrospinal fluid using OpenArray® platform for biomarker discovery
Authors: Liang Yu , Mouanoutoua Mokang , Shen Jianhe , Terashima Keita , Lau Ching 
Published: August 2012

Abstract
OpenArray® platform can be used for expression profiling using TaqMan® assays that enables higher throughput, less assay consumption, and lower sample input than other low-density assay platforms. One OpenArray® plate has 3,072 through-holes, each of which requires only 33nl of reaction volume. We combined the workflow of Megaplex™ RT primer pools and pre-amplification with TaqMan® MicroRNA assays on OpenArray® plates. Each OpenArray® plate provides the capacity of 3 samples, and each run can take up to 3 plates with a run time about 2.5 hours. Therefore, each typical work day can have 4 instrument runs generating expression data of 754 miRNAs from 36 samples. This platform is ideal to identify potential miRNA biomarkers within a short period of time by screening a large cohort of clinical specimens that might have limitations in sample size, contamination of PCR inhibitors, or low abundance of target miRNAs. We optimized the original workflow of Megaplex™ RT primer pools and pre-amplification before applying to the OpenArray® plates to ensure increasing detection sensitivity while maintaining the linearity in plasma samples. In a proof-of-principle investigation, we applied the new protocol to a small panel of plasma and cerebrospinal fluid (CSF) specimens derived from pediatric patients diagnosed with either germinoma or non-germinoma germ cell tumors of the brain, and benchmarked with another miRNA expression profiling platform, TaqMan® Array Card (TAC), and OpenArray® plates demonstrated good concordance with the TAC. Expression patterns of miRNAs detected in our CSF samples are largely consistent with previously published miRNAs differentially expressed in primary cancerous tissues between these two types of intracranial germ cell tumors. (For Research Use Only. Not for use in diagnostic procedures.)
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 3155. doi:1538-7445.AM2012-3155
Abstract 3158: MicroRNA signatures associate with pathogenesis and progression of osteosarcoma
Authors: Salah Zaidoun , Jones Kevin B. , Mare Sara Del , Aqeilan Rami 
Published: August 2012

Abstract
Osteosarcoma is the most common sarcoma in the skeleton, but remains elusive with regard to molecular etiology. MicroRNAs (miRNAs) have demonstrated far-reaching effects on the cellular biology of development as well as oncogenesis. Here we identify an miRNA signature reflecting the pathogenesis of conventional (osteoblastic/fibroblastic) osteosarcoma from surgically procured samples from human patients. The signature includes high expression of miR-181a, miR-181b, and miR-181c as well as reduced expression of miR-29b, miR-16, and miR-142-5p. Osteosarcomas of varied histology subtype co-localized on unsupervised hierarchical clustering, suggesting that the signature profile relates to oncogenesis itself, rather than simply a differentiation phenotype. We also demonstrated that miR-181b and miR-29b exhibit restricted expression to distinct cell populations in the tumor tissue. Further, higher expression of miR-27a and miR-181c* in pre-treatment biopsy samples characterized patients who developed clinical metastatic disease. In addition, higher expression of miR-451 and miR-15b in pre-treatment samples correlated with subsequent positive response to chemotherapy. In vitro functional validation of miR-16 and miR-27a confirmed opposing roles in human osteoblast and osteosarcoma cell lines. Furthermore, predicted target genes for miR-16 and miR-27a were confirmed as down-regulated by real-time PCR. Affymetrix array profiling of cDNAs from osteosarcoma specimens and controls were interrogated according to predicted targets of miR-16, miR142-5p, miR-29b, miR-181a/b, and miR-27a. This analysis revealed positive and negative correlations highlighting pathways of known importance to osteosarcoma, as well as novel genes. Thus, our findings establish miRNA signatures associated with pathogenesis, metastasis, and responsiveness to treatment in osteosarcoma.
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 3158. doi:1538-7445.AM2012-3158
Abstract 3207: Monitoring changes in NF-kB pathway regulation using highly sensitive multiplex bioluminescent reporter assays
Authors: Dobbs Megan , Hughes Douglas , Narahari Janaki , Choi Jae , Deshpande Atul , Webb Brian 
Published: August 2012

Abstract
Monitoring gene activation and repression in cellular signaling pathways requires specific tools. Reporter gene assays use a technique of attaching a genetic regulatory element, or promoter, to a gene encoding for a protein with enzymatic, chromophoric or fluorescent properties. Cloned into a plasmid vector, these sequences can be transfected or transduced into animal, plant or tissue culture cells. Reporter gene expression can be measured via bioluminescent, chemiluminescent or fluorescent output to quantify the element or protein activity in the pathway affecting the regulatory element. Bioluminescent output is formed through the catalysis of a substrate and the natural enzyme luciferase. Present in many terrestrial and marine metazoans, bioluminescence is a powerful instrument for mutational analysis of gene promoters, transcription factor dynamics, and protein-protein interactions. Our study confirmed the beneficial use of sensitive bioluminescent enzymes in reporter gene assays with wide dynamic ranges and flexibility in signal identification. Mutation of several of the luciferase genes resulted in spectral shifting and narrowing of the photon reaction products relative to the wavelength of the natural state. The mutations provide the ability to spectrally resolve the modified luciferases in a single reaction. In addition, multiplex assays relying on sequential addition of substrates can be achieved through manipulation of the enzyme-substrate reaction conditions. Marine copepod and ostracod luciferase enzymes are naturally secreted, and reporter assays using these genes can capture real-time or time course data by sampling the media of transfected cells or blood from transgenic animals and assaying for luciferase. The distinct advantage of these assays is that they are not destructive to the cells or animals. Our study used spectral separation using Gaussia/firefly and multiplex assays relying on sequential addition of substrates and Gaussia/Cypridina to monitor changes in NFkB activity in response to small-molecule agonists. Our results demonstrate the usefulness of dual-secreted luciferase assays for sensitive real-time monitoring of NFkB activity in media. Furthermore, these assays enable simultaneous detection of spectrally resolvable luciferases using filter-based detection.
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 3207. doi:1538-7445.AM2012-3207
Abstract 321: IL-6 in the tissue microenvironment plays a direct role in normal prostatic neoplastic transformation
Authors: Wu Jennifer D. , Liu Gang 
Published: August 2012

Abstract
Increased elevated levels of serum IL-6, the major inflammatory mediator, has been shown to be associated with development of progressive diseases in many types of cancers. We have previously shown that chronic exposure of immortalized benign prostate epithelial cells to paracrine IL-6 can induce tumorigenic transformation and further EMT (epithelial-mesochymal-transition)-associated metastasis in xenografts through stimulating autocrine IL-6 signaling and transactivating type I Insulin-like Growth Factor Receptor (IGF-IR). Whether IL-6, as a result of inflammation, in the tissue microenvironment can solely initiate tissue-specific de novo tumorigenesis is unclear. To address this question, we have generated a novel line of prostate-specific IL-6 transgenic mouse, in which human IL-6 (hIL-6) was expressed under the prostate-specific probasin promoter. We found that enforced expression of hIL-6 transgene in normal mouse prostate epithelium is sufficient to induce neoplastic transformation of the prostate gland, demonstrated by disappearance of base cell layer of the prostate gland and loss of E-cadherin and increased proliferation of the epithelium. Expression of the hIL-6 transgene also induced autocrine mouse IL-6 secretion in the prostate gland and sustained activation of STAT3 pathway in the epithelium and the stroma. Strikingly, IL-6 transgenic mice have increased peri-prostatic adipose tissue in which STAT3 was highly phosphorylated in comparison to the wild-type littermate. There is no significant increase in infiltration of T cells or B cells in the prostate of IL-6 transgenic animals. However, significantly increased infiltration (P&lt;0.01) of FoxP3+ lymphocytes was seen in the stroma of the prostate in the IL-6 transgenic animals in comparison to wild-type littermates. These data suggest that IL-6 as a pro-inflammatory cytokine alone can induce tissue-specific tumorigenesis through induction of autocrine IL-6 secretion and remodeling tissue microenvironment. Our finding suggests that inflammation may play a direct role beyond being the “stimuli” or “fuel” 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 321. doi:1538-7445.AM2012-321
Abstract 3214: Resveratrol potentiates glycolytic inhibitor-induced neuroblastoma cell death independent of sirt activity
Authors: Graham Regina M. , Puerta Nataly , Webster Keith A. , Vanni Steven 
Published: August 2012

Abstract
Introduction: Low survival rates and severe side effects of current neuroblastoma treatments underline the need for more effective less toxic therapies. Compared to normal cells, cancer cells demonstrate high glucose consumption even under aerobic conditions, therefore; targeting the glycolytic pathway in cancer cells as a potential therapy is currently an area of intense investigation. The polyphenolic compound resveratrol which can be found in grapes, peanuts, red wine and purple grape juice, has demonstrated antitumor activities both in vitro and in vivo. Here we investigate the potential of inhibiting glycolysis in combination with resveratrol treatment. Methods: To inhibit glycolysis we used the glucose analog, 2-Deoxy-D-glucose (2-DG). N-MYC amplified neuroblastoma cells (NB1691) were exposed to low dose 2-DG (0.5 and 2mM) and resveratrol 10uM. Cell viability was determined at 72hr using MTS assay. Western blotting was used to determine cell signaling pathways and apoptotic protein levels. The role of the NAD+ dependent HDAC sirtuin 1 (sirt1) was determined using 50uM of the inhibitor sirtinol. Results: While 2-DG did reduce cell viability (2mM:82+4.4% of control), the combination of 2-DG and resveratrol significantly reduced cell viability (20+2.2%) compared to resveratrol alone (69+2.8%). Western blot analysis indicated a robust activation of AKT with 2-DG with treatment, however this activation was blocked with 10uM resveratrol. Furthermore, western blot analysis revealed that this was caspase dependent cell death. PI3K/Akt inhibitors wortmannin and LY-294002 (10uM each) also reduced cell viability in combination with 2-DG to approximately 40% compared to non-treated, confirming the protective role of AKT signaling in response to 2-DG treatment. To determine if resveratrol, a well-known activator of sirt1, inhibited AKT via sirt1, cells were pretreated with sirtinol. Inhibiting sirt1 did not prevent resveratrol's inhibition of AKT. Conclusions: The oncoprotein AKT mediates signaling processes involved in evading apoptosis and promoting cell proliferation and recently has been linked to glycolysis. Resveratrol significantly increased 2-DG-induced cell death by inhibiting the 2-DG induced increase in AKT activity independent of sirt1. Here we show that combining resveratrol with 2-DG, two relatively non-toxic agents can significantly reduce neuroblastoma cell viability.
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 3214. doi:1538-7445.AM2012-3214