bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract LB-122: Single copy detection of KRAS mutations in colorectal cancer tissue and blood by qPCR using primers with enhanced analytical selectivity
Authors: Chupreta Sergey , Kelchner Vanessa , Kurihara Laurie , Makarov Vladimir 
Published: June 2012

Abstract
The development of highly sensitive genotyping assays that are suitable for clinical diagnostics opens new opportunities for the detection, assessment and management of cancer. Anticipated uses for these assays include profiling tissue biopsies and circulating tumor cells (CTCs), and detecting mutations in circulating cell-free nucleic acids. Swift Biosciences has developed myT™ Primers which have unique thermodynamic properties that make them highly sensitive to mismatch discrimination. myT™ Primer qPCR assays have been developed for the seven most frequent KRAS mutations in codons 12 and 13. These assays demonstrated single copy sensitivity for all seven KRAS mutations when used to genotype formalin fixed paraffin embedded (FFPE) specimens from Stage III colorectal tumors. When compared to a leading commercially available KRAS qPCR mutation test kit, myT™ Primer KRAS assays demonstrated several orders of improved specificity. A myT™ Primer assay was also developed for BRAF V600E/K mutations; results from FFPE melanoma specimens indicate high sensitivity and specificity. As a proof of concept for CTC detection, limiting dilutions of cells harboring the KRAS G12V mutation were added to whole human blood. Single copy detection of G12V was observed from an equivalent of a few microliters of blood. Because CTCs and cell-free cancer DNA can exist in blood at extremely low frequencies, we tested assay performance at even lower target concentrations. Mutant KRAS plasmid templates were titrated with wt KRAS plasmids such that the total KRAS copy number was 1×108. Under these conditions, the G12V-specific assay was able to detect the presence of ten mutant templates per 100 million (108) wt templates. The extreme selectivity of the myT™ Primer assays may make them especially useful for detection of mutations present at extremely low copy number and for genotyping difficult samples such as needle biopsies, CTCs and serum.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-122. doi:10.1158/1538-7445.AM2011-LB-122
Abstract LB-13: Calcitonin receptor-zonnula occludens-1 interaction is critical for calcitonin-induced destabilization of tight junctions and s prostate cancer metastasis
Authors: Shah Girish V. , Tamrakar Pratistha , Thomas Shibu , Lakshmikanthan Vijaybasker , Parajuli Parash , Sathyanarayanajois Seetharama , Bahouth Suleiman , Fanning Alan , Anderson James 
Published: June 2012

Abstract
Most primary prostate cancers (PCs) and PC cell lines express calcitonin (CT) and/or its receptor (CTR), and their co-expression positively correlates with tumor grade of primary PCs and growth/invasiveness of PC cell lines. Activation of the CT-CTR axis in non-invasive PC cells induces an invasive phenotype. In contrast, silencing of CT-CTR axis markedly reduces the tumorigenicity and metastatic capacity.
We have identified that cytoplasmic (C) tail of CTR contains a canonical class I type PSD-95, Discs-large, Zonnula Occludens-1 (PDZ)-binding motif, mutation of which does not affect its ability to bind its ligand or activate its G protein-coupled actions but abrogates CT-elicited destabilization of tight junctions or invasion. We then used CTR-C tail as a bait to identify its interactor by yeast two hybrid screening of prostate cDNA library. This revealed that CTR-C tail binds to intracellular membrane protein zonula occludens (ZO-1) through its PDZ domain. This interaction was confirmed using multiple approaches such as pull-down assays, overlay assays, co-immunoprecipitation and acceptor photobleaching FRET microscopy. The results demonstrate that CTR binds to PDZ3 domain of ZO-1 through its PDZ-binding motif. Mutation of either CTR-C PDZ-binding motif or ZO-1- PDZ3 domain abrogated actions of CT on tight junctions, invasion, orthotopic tumor growth and promotion of distant metastasis.
These results for the first time demonstrate PDZ-dependent interaction between CTR and ZO-1; and that this interaction is critical for CT-CTR axis-elicited prostate cancer progression. Considering overexpression of CT-CTR axis in malignant prostate, the results suggest that prevention of this interaction may serve as a new therapeutic possibility for the treatment of advanced prostate cancer.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-13. doi:10.1158/1538-7445.AM2011-LB-13
Abstract LB-137: YAP promotes breast cell proliferation and survival partially through stabilizing the KLF5 transcription factor
Authors: Zhi Xu , Zhao Dong , Zhou Zhongmei , Liu Rong , Chen Ceshi 
Published: June 2012

Abstract
The Yes-associated protein (YAP) is an oncoprotein in the Hippo tumor suppressor pathway that regulates tumorigenesis. Although YAP functions through its WW domains, the YAP WW domain binding partners have not been completely determined. In this study, we demonstrate that YAP functions partially through binding to KLF5, a transcription factor promoting breast cell proliferation and survival. YAP interacts with the KLF5 PY motif through its WW domains, prevents WWP1, a KLF5 E3 ubiquitin ligase, from binding to KLF5, and stabilizes KLF5 by decreasing the WWP1-mediated KLF5 ubiquitination and degradation. Over-expression of the wild type but not WW domain deficient YAP up-regulates the KLF5 protein levels and the mRNA expression levels of the KLF5 downstream target genes, including FGF-BP and ITGB2. In addition, knockdown of YAP decreases the expression levels of KLF5, FGF-BP, and ITGB2. Depletion of either YAP or KLF5 decreases breast cell proliferation and survival in MCF10A and SW527 breast cells. Importantly, stable knockdown of YAP or KLF5 significantly suppresses SW527 xenograft growth in mice. The YAP upstream kinase LATS1 suppresses the KLF5-FGF-BP signaling axis and cell growth through YAP. Finally, both YAP and KLF5 are co-expressed in estrogen receptor (ERα)-negative breast cell lines. These findings suggest that KLF5 is an important transcription factor partner for YAP and contributes to the Hippo tumor suppressor pathway. This study is supported by a grant from the American Cancer Society (RSG-08–199–01) and a grant from the Department of Defense (W81XWH-07–1–0191).
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-137. doi:10.1158/1538-7445.AM2011-LB-137
Abstract LB-139: A novel mechanism of tumor suppression by Piasy through NF-kappaB/RelA DNA binding repression and cytoplasmic translocation
Authors: Bridges Rebecca , Wang Yun , Wortham Aaron , Swanzey Emily , Stupor Zach , Lagowski James , Kulesz-Martin Molly , Liu Yuangang 
Published: June 2012

Abstract
NF-kappaB activation is commonly associated with malignancy. Constitutive NF-kappaB activation contributes to tumor growth, metastasis, and resistance to chemotherapy and radiation therapy. Activators of NF-kappaB in human cancer include viral oncogenes, inflammatory cytokines, and the loss of tumor suppressors like CYLD. Loss of the protein inhibitor of signal transducer and activator of transcription family member Piasy is associated with malignant progression in chronic myelogenous leukemia and myelodysplastic syndrome, suggesting its role as a tumor suppressor. In this study, we provide evidence for a novel mechanism for NF-kappaB activation in human cancer through deficiency of Piasy. Piasy repressed NF-kappaB luciferase activity in response to TNFalpha, constitutively active IKK2, or overexpressed RelA. Piasy protein directly bound to RelA, through the C terminus of Piasy and the DNA binding domain of RelA, distinguishing it from other Pias family proteins that bind to RelA. Piasy inhibited DNA binding of RelA in cells, as detected by DNA affinity immunoblotting and chromatin immunoprecipitation. However, the DNA binding activity of purified RelA protein was not affected by purified Piasy protein, suggesting that Piasy indirectly represses RelA DNA binding in vivo. Analysis of cellular localization of Piasy and RelA proteins revealed that Piasy colocalized with RelA in the cytoplasm, correlated with enhanced RelA cytoplasmic translocation. In vivo consequences of Piasy depletion were shown in a mouse model of psoriasis-like epidermal hyperplasia in mice, and in human squamous cell carcinomas (SCC) of the skin. Piasy null mice topically treated with NF-kappaB activator imiquimod exhibited increased nuclear RelA in epidermis compared to wild type controls. Piasy levels in human skin SCC were inversely associated with nuclear RelA. These findings suggest a novel mechanism for Piasy as a tumor suppressor by the inhibition of NF-kappaB oncogenic pathways in human cancers. (This work was supported by NIH 5R01CA098577, and 5R01AR55651)
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-139. doi:10.1158/1538-7445.AM2011-LB-139
Abstract LB-140: Menin-mediated regulation of pancreatic β-cell differentiation factors
Authors: Shi Kerong , Parekh Vaishali , Roy Swarnava , Agarwal Sunita K. 
Published: June 2012

Abstract
Pancreatic endocrine tumors occur sporadically or in familial multiple tumor syndromes such as MEN1 (Multiple Endocrine Neoplasia type 1). Biallelic loss/inactivation of the MEN1 tumor suppressor gene is only found in 20–30% of sporadic pancreatic endocrine tumors. Therefore, identification of other molecular and genetic causes of pancreatic endocrine tumorigenesis is an area of intense investigation. Because MEN1 is the main gene reported so far that affects pancreatic endocrine tumors, analyzing downstream effects of menin (MEN1 gene encoded protein) could uncover other candidate genes and pathways responsible for pancreatic endocrine tumorigenesis. During the process of endocrine pancreas development, a cascade of transcription factors under tight sequential- and temporal-regulated expression ensures normal organogenesis. Alteration of such developmental and differentiation factors could potentially lead to tumorigenesis.
To study cellular transformation related to menin loss in the pancreatic β-cell, we used the mouse pancreatic β-cell line MIN6 as a model system. RNA and protein were isolated from MIN6 cells after menin knockdown or after menin over-expression as well as from their respective controls. Twelve transcription factors associated with β-cell differentiation were analyzed using QRT-PCR and western blot: Hlxb9, Hnf3β, Isl-1, MafA, MafB, NeuroD1, Ngn3, Nkx2.2, Nkx6.1, Pax4, Pax6, and Pdx1.
Three different sources of menin siRNAs and four different transfection reagents were tested in time course knockdown experiments. Significant menin knockdown in MIN6 cells was observed with 74.3% ± 15.03% (n=6) knockdown efficiency at 72 hours post-transfection. For transient menin over-expression in MIN6 cells, the transfection efficiency was close to 60% (based on GFP plasmid transfection). Endogenous expression of all factors, except Ngn3, was detectable in MIN6 cells. None of the eleven transcription factors showed significantly altered RNA expression upon menin knockdown or upon menin over-expression. Western blot analyses showed 4–5 factors that were up-regulated upon menin knockdown, and down-regulated upon menin over-expression. Evolutionarily conserved promoter regions of Hlxb9, Isl-1, Nkx6.1, or Pax4 were not regulated upon menin knockdown or upon menin over-expression. These data indicate that menin could regulate some β-cell differentiation factors in a post-transcriptional manner. Mechanism of menin-mediated post-transcriptional regulation and downstream consequences on β-cell transformation and tumorigenesis is under investigation.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-140. doi:10.1158/1538-7445.AM2011-LB-140
Abstract LB-142: Regulation of MYCN stability by reactive oxygen species in neuroblastoma
Authors: Ikegaki Naohiko , Regan Paul L. , Hicks Sakeenah L. , Maloney Nolan , Tang Xao X. 
Published: June 2012

Abstract
Neuroblastoma (NB) is the most common extracranial solid tumor in children. Amplification of the MYCN proto-oncogene is associated with older age, rapid tumor progression, and the worst outcome of this disease. As MYCN amplification leads to its over-expression, high-level expression of MYCN is thought to cause an aggressive behavior of MYCN amplified tumors. In fact, the forced reduction of MYCN protein expression by siRNA results in growth suppression and apoptosis of NB cells with MYCN amplification. We have identified several compounds that can rapidly destabilize the MYCN protein (within a few hours), in NB cells and cause growth suppression. The compounds include FCCP, OSU-03012, and Salinomycin. Our recent data suggest that a common effect of the above compounds appears to be the inhibition of mitochondrial functions.
Moreover, ascorbic acid, an anti-oxidant, abolishes the effect of these compounds on MYCN protein stability. It is known that inhibition of mitochondrial oxidative phosphorylation increases the production of reactive oxygen species (ROS), which include superoxide (O2-), hydroxyl radical (•OH), and hydrogen peroxide (H2O2). Anti-oxidants are also known to quench ROS. Preliminary data showed that ROS was detected when the IMR5 NB cells were treated with OSU-03012 for three hours. In addition, we have found that forced over-expression of pVHL, an E3 ubiquitin ligase, in NB cell lines results in reduction of MYCN protein levels. We are currently testing whether ROS regulates MYCN stability by hydroxylation of proline residues on MYCN, and whether pVHL recognizes the ROS-modified MYCN, which is then subjected to rapid proteasome-dependent degradation.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-142. doi:10.1158/1538-7445.AM2011-LB-142
Abstract LB-213: Acheron augments triple negative breast cancer stem-like cells and exacerbates chemotherapeutic and radiation resistance
Authors: Scully Steve J. , Compton Shannon , Francescone Ralph A. , Schwartz Lawrence , Yan Wei , Shao Rong 
Published: June 2012

Abstract
Basal-like breast cancers that are negative for clinically important receptors (estrogen, progesterone, and Her2/neu), carry the worst prognosis for patients that are diagnosed with breast cancer. Acheron is a novel protein that was recently demonstrated to be highly expressed in triple-negative basal-like breast cancers. Previous work in our lab has demonstrated that breast cancer cell lines that are engineered to express ectopic Acheron are more invasive, have higher levels of MMP-9 and VEGF, and develop increased tumor burden in vivo in xenografted SCID/beige mice.
In order to better understand the role Acheron may play in these basal-like breast cancers, the triple negative breast cancer cell lines MDA 231 and MDA 435 were engineered to express ectopic Acheron. Engineered cells were cultured in serum free media with EGF and FGF to cultivate them as mammary cancer stem-like cells. Relative to empty vector control cells, Acheron-expressing MDA 435 cells demonstrated a greater ability to form acini-like structures in suspension as well as support anchorage independent growth.
Cells that were cultivated in basal media with serum were also analyzed for cell adhesion and expression of the laminin adhesion protein CD29 (Beta-I Integrin). Expression of Acheron in either MDA 231 and 435 cells lead to a marked reduction in substrate adhesion on laminin-coated plates relative to controls. Consistent with this observation, Acheron-expressing cells also expressed reduced levels of CD29.
FACS analysis for the apoptosis markers Annexin-V binding and propidium iodide exclusion demonstrated that Acheron-expressing cells were 2–3 times less sensitive to the same concentration of the chemotherapeutic drugs: Doxorubicin and docetaxel, and to a lesser extent to ionizing radiation (1.6 fold).
In summary, we have extended the analysis of Acheron action in triple-negative basal-like breast cancer cells. We have found that these cells that express ectopic Acheron are more resistant to chemotherapeutic drugs and radiation and have a greater capacity to grow in an anchorage-independent manner. These data support the hypothesis that Acheron expression in triple-negative breast cancers may confer properties that would be anticipated to result in a poorer prognosis. Therefore, Acheron deserves attention and further investigation as a new cancer target in triple-negative basal-like breast cancer.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-213. doi:10.1158/1538-7445.AM2011-LB-213
Abstract LB-220: TMEM16A, a novel calcium-activated chloride channel, modulates tumor proliferation via MAPK and Cyclin-D1 signaling
Authors: Shiwarski Daniel , Bertrand Carol , Egloff Ann Marie , Huang Xin , Seethala Raja , Grandis Jennifer , Gollin Susanne , Duvvuri Umamaheswar 
Published: June 2012

Abstract
TMEM16A, a calcium-activated chloride channel, was recently isolated from oral cavity cancer cells. TMEM16A is amplified in head and neck cancers and overexpressed in gastrointestinal stromal tumors (GIST's); however, the role that TMEM16A plays in promoting tumor growth remains unclear. The goal of this research was to establish the biologic function of TMEM16A in the context of epithelial malignancies. We hypothesized that TMEM16A contributes to tumor cell survival and may be a useful prognostic factor for patients with squamous cell carcinoma of the head and neck (SCCHN). Knockdown and overexpressing SCCHN and T24 cell lines were transduced with lentiviral particles encoding RNAi and engineered to stably overexpress TMEM16A. For functional verification, electrophysiologic assays were performed to establish that the altered TMEM16A expression was modulating the chloride current. To elucidate a mechanism of TMEM16A's proliferative effects, immunoblotting and pharmacologic inhibition of potential downstream targets were performed. Our results demonstrate that the levels of TMEM16A expression modulates proliferation in vitro and in vivo, and that stable overexpression of TMEM16A increases resistance to cisplatin therapy as compared to cancer cell lines expressing endogenous TMEM16A. Specificity of these phenotypic results were confirmed using genetic rescue experiments. Next, we identified that the MAPK/Cyclin-D1 axis was a critical signaling pathway in TMEM16A-induced proliferation, and that pMAPK/MAPK and Cyclin-D1 levels paralleled expression of TMEM16A. Finally, using pharmacologic agents niflumic acid or NPPB to inhibit the TMEM16A chloride channel activity in combination with a MAPK inhibitor (U0126) demonstrated additive effects significantly decreasing cancer cell proliferation in vitro. In summary, our results provide novel insights into the biologic function of TMEM16A and its role in tumor cell proliferation. This research has enhanced our understanding about the pathogenesis of SCCHN tumors, and can potentially be translated to many other cancers.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-220. doi:10.1158/1538-7445.AM2011-LB-220
Abstract LB-229: Arsenic trioxide, ascorbic acid and disulfiram (AAA) arrests pancreatic cancer development in a transgenic mouse model with p16 knockout and activated Kras
Authors: Dinnen Richard D. , Qiu Wanglong , Su Gloria H. , Fine Robert L. 
Published: June 2012

Abstract
Previously, we found that four human pancreatic cancer (PC) cell lines underwent robust cell death associated with dramatic ATP decline and ROS accumulation when treated with clinically achievable concentrations of arsenic trioxide, ascorbic acid and disulfiram (AAA). Early apoptosis was shifted to aponecrosis, and dependent on the voltage-dependent anion channel (VDAC), since shRNA directed against VDAC-1 abrogated the ROS accumulation and blocked cell death. In this study, assessment of AAA was performed in Pdx1-Cre transgenic mice with a conditional p16 knockout (p16flox/flox), and oncogenic KrasG12D. The KrasG12D, p16flox/flox; LSL-KrasG12D; Pdx1-Cre mice develop the full spectrum of pancreatic intraepithelial neoplasia (mPanIN) lesions, pancreatic ductal adenocarcinoma, and hepatic and peritoneal metastases. Over 95% of human PC have p16 deficiency and Kras mutation. Eight week old mice were treated for 8 weeks, 3 times/week with AAA, sacrificed, and the development of PC assessed. Little or no toxicity of the mice to the AAA treatment was observed. Frank PC was zero in AAA-treated mice and 31% in controls. The mean number of PanIN-3 lesions/mouse declined from 14.1 in controls to 1.58 in AAA-treated mice (p=0.008). The mean number of PanIN-2 lesions declined from 5.18 in controls to 1.67 in AAA treated mice (p=0.06). The mean number of PanIN-1 lesions in control mice were 14.2 and in AAA-treated mice were 16.0, p=0.51 indicating no difference in PanIN-1 occurrence. Preliminary results from immunohistochemical staining of the PanIN lesions for VDAC, indicated that PanIN-1 lesions have low, and PanIN-2 and 3 lesions have high, VDAC expression. This result parallels the in vitro human cell line data showing VDAC expression is critical to AAA sensitivity, and suggests a similar mechanism of action in vivo as in vitro. Although collection of data is ongoing, this is the first treatment that has been shown to arrest PanIN and tumor development in any a genetically-engineered mouse model for PC. The data suggests AAA either arrests the development of pre-cancerous lesions after the PanIN-1 stage, or the PanIN-2 and PanIN-3 lesions are destroyed by AAA in a VDAC-dependent manner.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-229. doi:10.1158/1538-7445.AM2011-LB-229