bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 2866: Sodium channel γENaC mediates salt induced inflammatory stress in cancer cells
Authors: Amara Suneetha , Tiriveedhi Venkataswarup 
Published: July 2016

Abstract
Chronic inflammation is known to play a critical role in the development of cancer. Recent evidence suggests that high salt in the tissue microenvironment induces chronic inflammatory milieu. Further, high salt intake has been correlated with higher incidence of cardiovascular diseases and cancers. In this report, using breast cancer cell lines, MDA-MB-231 (highly invasive), MCF-7 (poorly invasive), and a normalized breast epithelial cell line, MCF10A, we determined the molecular basis of the potential inflammatory effect of high sodium chloride (NaCl) in the extracellular environment. Dose response studies demonstrated a significantly reduced cell viability (&gt;50%) with external NaCl concentration above 0.25 M. Combined treatment of high NaCl (0.15 M) with sub-effective interleukin (IL)-17 (0.1 nM) induced reactive nitrogen (RNS) and oxygen (ROS) species release along with upregulation of the enzymes iNOS, SOCS and SOD1. Similar effect was not observed with equi-molar mannitol. Western-blot and qRT-PCR analysis demonstrated an elevated expression of an inflammation-specific epithelial sodium channel, γENaC, in MDA-MB-231 (7.1±1.4 fold, p&lt;0.05), MCF-7 (3.9±0.8 fold, p&lt;0.05) and MCF10A (2.2±0.4 fold, p&lt;0.05) cell lines following combined treatment of high NaCl with sub-effective IL-17. This was significantly abrogated by lipid raft depletion and ERK-1/2 knock-down. These culture conditions have also induced cellular expression of cytokines, IL-6 (3 fold), TNFα (6.2 fold); chemokines, CCL5 (2.3 fold), CXCL-12 (2 fold), MIP-1δ (2.7 fold); and angiogenic growth factor, VEGF (2.5 fold). Taken together, these data suggest that high NaCl in the cellular microenvironment induces a γENaC mediated chronic inflammatory response with a potential pro-carcinogenic effect.
Citation Format: Suneetha Amara, Venkataswarup Tiriveedhi. Sodium channel γENaC mediates salt induced inflammatory stress in cancer cells. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2866.
Abstract 2869: FLASH-dependent post-translational regulation of the EMT activator ZEB1
Authors: Abshire Camille , Dragoi Ana Maria 
Published: July 2016

Abstract
Understanding the mechanisms of metastasis initiation and tumor persistence despite aggressive treatments is paramount to developing new viable therapeutic targets. Critical to tumor progression is initiation of epithelial-to-mesenchymal transition (EMT), a cellular process characterized by loss of E-cadherin, a critical cell-cell adhesion molecule, gain of migratory properties and chemoresistance. Transcriptional repression of E-cadherin and other genes responsible for maintaining the epithelial phenotype by ZEB1/ZEB2 family is one of the most significant events occurring at the interface between non-metastatic/metastatic cancer and chemo-sensitive/chemo-resistant cancer. Using a novel interdisciplinary high-throughput screen methodology we uncovered novel regulators of E-cadherin. Caspase 8 - associated protein 2 (CASP8AP2 or FLASH) a protein involved in apoptosis and transcriptional control was identified through this screen and we discovered that FLASH depletion resulted in increased E-cadherin levels at the plasma membrane through loss of ZEB1 expression. Subsequent examination of ZEB1 protein expression revealed that ZEB1 is rapidly degraded in FLASH-depleted cells. This process is blocked by proteasome inhibitors suggesting that post-translational modifications (PTMs) and in particular ubiquitination of ZEB1 is responsible for its fast turnover and decreased protein expression in the absence of FLASH. Indeed, ubiquitinated ZEB1 was detected in FLASH-depleted cells in the presence of proteasome inhibitors. Although ZEB1 is degraded through the ubiquitin pathway, its rapid turnover in the absence of FLASH suggests that protective mechanisms are in place to delay and/or prevent this process. Even more, we determined that the atypical ubiquitin E3 ligase complex Skp1-Pam-Fbxo45 is involved in ZEB1 degradation in the absence of FLASH and is upregulated in cells depleted for FLASH. Altogether our data suggests that FLASH can stabilizes ZEB1 indirectly by transcriptional repression of the ubiquitin ligase responsible for its degradation and directly by binding to ZEB1 and masking the recognition site of the ubiquitin ligase. Even more, absence of FLASH reprograms the cells into a more epithelial-like phenotype which is less invasive and is resistant to TGFβ-induced loss of E-cadherin. Not only is FLASH-dependent post-translational regulation of ZEB1 novel, but is also conserved in multiple cells lines from various tissues, making this mechanism an original therapeutic target for cancer metastasis.
Citation Format: Camille Abshire, Ana Maria Dragoi. FLASH-dependent post-translational regulation of the EMT activator ZEB1. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2869.
Abstract 2870: Serine/arginine rich proteins show different distribution patterns in stratum of oral epidermoid carcinoma
Authors: Ceja-Lopez Cynthia J. , Lara-Martínez Reyna , Segura-Valdés Lourdes , Ruíz Godoy Rivera Luz , Ambrosio Hernandez Javier A. , Guadarrama-Orozco Jorge Alberto , Ruíz-García Erika , Jiménez García Luis Felipe , Meneses-García Abelardo 
Published: July 2016

Abstract
Serine/arginine proteins increased their activity during the alternative splicing in cell nucleus. The nuclear speckles, also known as interchromatin granule clusters, are enriched with SR splicing factors and are implicated in gene expression. Nuclear speckle formation is developmentally regulated; in certain cases phosphorylated SR proteins are absent from the nucleus and are instead localized at granular structures in the cytoplasm. SR proteins are related with splicing during different types of cancers and metástasis. We search for phosphorylated SR proteins and the nuclear pattern and speckle formation in human oral carcinomas.
Methods: To address nuclear pattern and speckle formation, we performed a series of immunostaining experiments, with SC35 antibody for phosphorylated srsf2 on sections derived from oral epidermoid tumor from 33 patients. The images were obtained with a microscope (Leica TCS SPE/CTR 4000). For imaging cytometry analyses, a Leica LAS AF Lite software (Leica Microsystems) was used to capture images automatically; images and recognition of nuclear speckles were quantitatively analyzed using analysis software (ImageJ). ANOVA analysis was performed for the cellular and staining correlation.
Results: There are significant differences in morphology and distribution of intranuclear mottled pattern of SR proteins in each tissue layer between oral epithelial without neoplasia and oral squamous cell carcinoma (P &lt; 0.0001). We also found that number, size and pattern (p &lt;0.05) is different between the stratum layers of the oral squamous cells, showing more transcriptional activity in the granulosum stratum related with a decreased in nucleus numbers. In oral squamous cells the pattern changes become homogeneous and similar to the basal layer of normal tissue.
We found that is in the granulosum stratum of oral squamous carcinomas where the SR proteins shows more transcriptional activity. This finding could be related with the increased metabolism of this layer.
Citation Format: Cynthia J. Ceja-Lopez, Reyna Lara-Martínez, Lourdes Segura-Valdés, Luz Ruíz Godoy Rivera, Javier A. Ambrosio Hernandez, Jorge Alberto Guadarrama-Orozco, Erika Ruíz-García, Luis Felipe Jiménez García, Abelardo Meneses-García. Serine/arginine rich proteins show different distribution patterns in stratum of oral epidermoid carcinoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2870.
Abstract 2877: Effects of metabolic stressors on cellular differentiation
Authors: Schvartzman Juan Manuel , Thompson Craig B. 
Published: July 2016

Abstract
Poor differentiation (the divergence of tumor cell morphology from the cellular morphology of the normal tissue of origin) is associated with unfavorable responses to chemotherapy and poor clinical outcomes in solid tumors. Recurrent somatic mutations in the genes encoding for the metabolic enzymes isocitrate dehydrogenase (IDH) 1 and 2 have recently been identified in patients with acute myeloid leukemias, chrondrosarcomas, cholangiocarcinomas and glioblastomas. Point mutations in IDH lead to production of 2-hydroxyglutarate (2HG). 2HG acts as a potent inhibitor of enzymes involved in the removal of chromatin marks associated with transcriptional repression (DNA and Histone H3K9 methylation). Excess 2HG leads to impaired differentiation but very little is known about the underlying mechanisms. Here we use a well established model of cellular differentiation, the differentiation of 10T1/2 fibroblasts into myofibers by the lineage-specifying transcription factor MyoD to study the mechanisms by which IDH mutations impair differentiation. We show that 5-azacytidine induced differentiation of 10T1/2 cells is impaired when mutant IDH2 is expressed and that IDH2 mutations impair the ability of MyoD expression to reprogram 10T1/2 cells into myofibers. We propose that the metabolic landscape of undifferentiated cells determines the ability of the lineage determining factor MyoD to trigger differentiation and that the undifferentiated state of cancer cells is dependent on their unique metabolic profile. Further work aims to a) define the effects of mutant IDH1/2 on the ability of MyoD to program muscle differentiation, b) determine the nature of the tumorigenic effects of IDH1/2 mutations in 10T1/2 cells and c) identify molecular pathways that can overcome differentiation blocks upon activation. Insight from these experiments will outline the mechanism(s) by which metabolic alterations, in the form of 2HG, impair differentiation and drive tumorigenesis. They will also serve as a platform for the identification of metabolic alterations that trigger differentiation. These mechanisms could prove to be novel and readily targetable processes in an array of poorly differentiated and aggressive cancer subtypes where other pharmacological approaches have failed.
Citation Format: Juan Manuel Schvartzman, Craig B. Thompson. Effects of metabolic stressors on cellular differentiation. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2877.
Abstract 2879: PIGU modulates radioactive iodine uptake in differentiated thyroid cancers
Authors: AMIT MORAN , Na’ara shorook , Charas Tomer , gil. Ziv 
Published: July 2016

Abstract
Radioactive iodine (RAI) treatment has a significant role in the management of differentiated thyroid carcinoma after surgical treatment. Although there is significant variability between patients, in response to RAI treatment, the mechanism responsible for this phenomenon is unknown. PIGU is an endoplasmic reticulum protein and a component of the GPIT complex, which plays a crucial role in post-translational modification of proteins such as sodium iodide symporter (NIS). Immunohistochemical analysis revealed significantly lower expression of PIGU in papillary carcinomas compared to matched normal thyroid tissue (P&lt;0.001). To study the role of PIGU in RAI uptake of differentiated thyroid cancer, we stably expressed PIGU and mock plasmid in the papillary cancer cell line (K1-PIGU and K1, respectively). Immunofluorescence analysis showed a significant increase in perinuclear expression of PIGU in the K1-PIGU cancer cells compared to controls. Similarly, flow cytometry analysis showed a significant increase in membranous expression of NIS in PIGU transfected cancer cells compared to controls. There were no significant differences in proliferation, invasion or motility between K1-PIGU and K1 cell lines. PIGU expression significantly increased I125 uptake compared to controls both in vitro (1.13 and 1030 ppm, respectively, p&lt; .001) and in vivo (153 and 1.2×105 ppm, respectively, p&lt; .001). Mitogen-activated protein kinase inhibition showed clinically meaningful increases in iodine uptake and retention in a subgroup of patients with thyroid cancer that is refractory to radioiodine. To address the mechanism underlying PIGU down regulation in RAI resistant tumors, we treated K1 and KI-PIGU cells with U0126 MEK inhibitor. This inhibition resulted in a significant increase in PIGU expression and I125 uptake in vitro. Taken together, our results revealed a pivotal role for PIGU in RAI sensitivity of thyroid cancer cells, by modulation of NIS expression.
Citation Format: MORAN AMIT, shorook Na’ara, Tomer Charas, Ziv gil. PIGU modulates radioactive iodine uptake in differentiated thyroid cancers. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2879.
Abstract 289: Melphalan and XPO1 inhibition are synergistic in pre-clinical models of multiple myeloma
Authors: Turner Joel G. , Dawson Jana L. , Sullivan Daniel M. 
Published: July 2016

Abstract
Introduction:
Multiple myeloma (MM) accounts for about 10% of all hematologic malignancies, with estimated numbers of new cases and deaths for 2015 in the US at 26,850 and 11,240, respectively. Significant increases in response/survival have been seen over the past several years; however, MM remains incurable and patients ultimately die from progressive disease refractory to anti-myeloma therapy. There is clearly a need for additional effective therapies.
Materials and Methods:
Human MM parental and melphalan (MEL) resistant cell lines were treated with XPO1 inhibitors (XPO1i) KPT330 or KPT8602 +/- MEL and assayed for apoptosis and viability by flow cytometry. Treated cells were assayed for DNA damage by comet assay and phospho-H2AX protein expression. XPO1/MEL treated cells were assayed for p53, NFkB, IKKα, FANCF, and FANCL by Western blot. Cells from patients with newly diagnosed or relapsed/refractory MM treated with XPO1i/MEL were assayed for apoptosis. NOD/SCID-gamma mice with MM tumors were treated with XPO1i/MEL and assayed for tumor growth, survival, and toxicity.
Results:
MM cell viability was decreased synergistically and apoptosis increased by XPO1I/MEL treatment in all MM cell lines tested. XPO1/MEL drug combination significantly increased DNA damage when compared to either MEL or XPO1 alone, as shown by comet assay and increased phospho-H2AX expression. Western blot showed that XPO1i treatment of MM cells increased p53 and decreased NFkB, IKKα, FANCF, and FANCL. MEL-resistant MM cell lines were found to be sensitized by XPO1i to MEL as shown by apoptosis assay (20-fold). CD138+/light chain+ MM cells from newly diagnosed and relapsed/refractory MM patients were also sensitized (5-10 fold) by XPO1i to MEL (apoptosis assay). NOD/SCID-gamma mice challenged with MM tumors demonstrated a strong synergistic antitumor effect in XPO1i/MEL-treated mice, with increased survival and no significant toxicity.
Conclusions:
XPO1i's improved the response of human MM cell lines and patient MM cells to MEL in vitro and ex vivo. It is possible that this synergy may be due to increased nuclear p53 in combination with decreased NFkB and IKKα and decreased DNA repair proteins FANCL and FANCF reversing MEL resistance by the Fanconi Anemia/BRCA pathway. These preliminary data suggest that XPO1i's augment MEL-induced DNA damage and may also block the repair of the DNA damage—both of which could result in synergistic cell kill. Combination therapies using XPO1i, especially the clinical compounds KPT330 (selinexor) and KPT8602 +/- MEL, may significantly improve the treatment outcomes of MM.
Citation Format: Joel G. Turner, Jana L. Dawson, Daniel M. Sullivan. Melphalan and XPO1 inhibition are synergistic in pre-clinical models of multiple myeloma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 289.
Abstract 2897: Recurrent transcriptome alterations across multiple cancer types
Authors: Kaczkowski Bogumil , Tanaka Yuji , Kawaji Hideya , Sandelin Albin , Andersson Robin , Itoh Masayoshi , Lassmann Timo , Hayashizaki Yoshihide , Carninci Piero , Forrest Alistair R.R. ,   FANTOM5 Consortium
Published: July 2016

Abstract
Genes that are frequently deregulated in cancer are clinically attractive as both potential pan-cancer diagnostic markers and therapeutic targets. Here we compared Cap Analysis of Gene Expression (CAGE) profiles from 225 cancer cell lines and 339 corresponding primary cell samples to identify transcripts that are recurrently deregulated in a broad range of cancer types. CAGE is a 5’ sequence tag technology that globally determines transcription start sites (TSS) in the genome and their expression levels. This allowed us to assess novel aspects of the cancer transcriptome.
First, we identified (at promoter resolution) hundreds of protein coding and long non-coding transcripts that are commonly de-regulated in cancer cell lines. Next, we showed that promoters that overlap repetitive elements (especially SINE/Alu and LTR/ERV1 elements) are often upregulated in cancer. In particular, a specific repeat family, REP522 (largely palindromic, unclassified interspersed repeat of ∼1.8Kb in size), was strongly enriched for the most up-regulated promoters. To our knowledge this is the first report implicating REP522 activation in cancer. Then, taking advantage of the fact that CAGE data can be used to estimate the activity of enhancers from balanced bidirectional transcription we identified 90 enhancer RNA producing regions that are recurrently activated in cancer cell lines. With ENCODE ChIA-PET data, we linked 16 of the cancer-activated enhancers to promoters of known cancer related genes.
Finally, to confirm that our results are relevant to clinical tumors we performed complementary analysis in RNA-seq data from 4,055 tumors and 563 normal tissues profiled by The Cancer Genome Atlas (TCGA) and we identified a core set of pan-cancer biomarkers (of both coding and non-coding transcripts) that are recurrently perturbed in both the FANTOM5 and TCGA datasets.
In summary, our extensive transcriptome analysis identified a comprehensive set of candidate biomarkers with pan-cancer potential, and extended the perspective of enhancers and repetitive elements that are recurrently activated during carcinogenesis.
Citation Format: Bogumil Kaczkowski, Yuji Tanaka, Hideya Kawaji, Albin Sandelin, Robin Andersson, Masayoshi Itoh, Timo Lassmann, Yoshihide Hayashizaki, Piero Carninci, Alistair R.R. Forrest, FANTOM5 Consortium. Recurrent transcriptome alterations across multiple cancer types. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2897.
Abstract 2899: Selinexor inhibits NF-κB activity by sequestering IkB-a in the nucleus and blocking IkB-a degradation
Authors: Kashyap Trinayan , Argueta Christian , Klebanov Boris , Unger TJ , Link Benjamin , Werman Maxwell , Lee Margaret , Shacham Sharon , Landesman Yosef 
Published: July 2016

Abstract
Background: Selinexor is a small-molecule therapeutic that inhibits XPO1 mediated nuclear export, resulting in nuclear accumulation of tumor suppressor proteins (TSPs) and subsequently cancer cell death while sparing normal cells. It has been previously demonstrated that the inhibitor of NFκB, IκB-, is localized to the cytoplasm by XPO1 in several cancer cell lines and that treatment of cancer cells with selinexor reduces NF-κB transcriptional activity. The mechanism of NF-κB inhibition by selinexor, however, is not fully understood. We hypothesized that nuclear retention of IκB- and down-regulation of IκB- kinase (IKK) in response to selinexor treatment would inhibit NF-κB transcriptional activity.
Methods: U2OS (Osteosarcoma) cells were treated with selinexor in the presence or absence of TNF stimulation and whole protein lysates were analyzed by Western blotting. IκB- localization was evaluated by immunofluorescence microscopy and NF-κB transcriptional activity by ELISA (Thermo Scientific).
Results: TNFα induced the phosphorylation of NF-κB p65 subunit on serine 536 and IκB- on serine 32/36 through IKK. This resulted in the dissociation of IκB-α from NF-κB and led to IκB-α degradation via the 26S proteasome. Free NF-κB could now migrate into the nucleus and initiate transcriptional activation supporting tumorigenicity and inflammation. The IKK kinase is a complex made of two kinases (IKKα and IKKβ) and one regulatory subunit, NEMO/IKKγ. We found that selinexor treatment blocked IKK activity through the down regulation of the IKK (IKKγ) gamma subunit protein levels. This inhibition was dose dependent and prevented IκB- phosphorylation, thereby protecting IκB- from degradation. The protection of intact IκB- from degradation and its forced nuclear accumulation through XPO1 inhibition enabled inhibition of NF-κB transcriptional activity even in the presence of TNFα. Selinexor did not alter the protein levels of IKK or IKK.
Conclusions: Selinexor blocks TNFα induced degradation of IκB- by reducing the levels of IKK. In addition, selinexor increased nuclear IκB- levels through the inhibition of nuclear export. This blocked NF-κB activity and enhanced cancer cell death. We are currently investigating the beneficial effects of combining selinexor with proteasome inhibitors, which are known to prevent IκB- degradation.
Citation Format: Trinayan Kashyap, Christian Argueta, Boris Klebanov, TJ Unger, Benjamin Link, Maxwell Werman, Margaret Lee, Sharon Shacham, Yosef Landesman. Selinexor inhibits NF-κB activity by sequestering IkB-a in the nucleus and blocking IkB-a degradation. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2899.