bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 1825: The inhibitory effect of long non-coding RNA MALAT1 on Smads signaling pathway.
Authors: Zhang Jinqiang , Han Chang , Wu Tong 
Published: August 2014

Abstract
Long non-coding RNA (lncRNA) is arbitrarily defined as RNA longer than 200 bases that does not encode mRNA, rRNA or tRNA. LncRNAs are involved in almost every aspect of cellular and molecular biology. In most cases, lncRNAs assemble with proteins as RNA-protein complexes to execute their functions. We hypothesize some important molecular in critical signaling pathway, such as Smad2 and Smad3 in TGF-β/Activin pathway, might have their lncRAN partner/partners to fine regulate their function. Based on the hypothesis, we carried on little scale screen and finally identified lncRAN MALAT1 can exclusively bind to pSmad2 and pSmad3 in hepatocellular carcinoma cell lines Hep3B, SK-Hep1 and cholangiocarcinoma cell line CCLP1 by methods of RNA Immunoprecipitation (RIP). After quantifying by qRT-PCR, we found that MALAT1 enriched by anti-pSmad2/3 antbodies is &gt;15 times more than that by IgG control. Co-localization of MALAT1 and pSmad2 and 3 in nuclear speckles of Hep3B cells were also proved by methods of combining FISH and immunofluorescence staining. Further bioinformatics analysis and immunoprecipitation reveal that pSmad2/3 binding to MALAT1 through SET domain containing 2 (SETD2) indirectly. Accordingly, if SETD2 was knockdown by siRNA, the binding between pSmad2/3 with MALAT1 was proved decreasing dramatically by methods of RIP. We also found that MALAT1 knockdown by shRNA or antisense oligonucleotide will increase the Smad2/3 signaling activity significantly, which was proved by Smad2/3 luciferase reporter constructs, DNA pull-down using synthesized oligonucleotide containing consensus sequence of Smads binding elements (SBE) and enhanced inhibition effect of TGF-β on the proliferation of Hep3B and CCLP1 cells. Next, Protein Phosphatase 1A (PPMA1), which act as a Smad phosphatase to terminate pSmad2/3 activity, was proved to bind to MALAT1 and MALAT1 knockdown decreased the binding of pSmad2/3 to PPM1A. Therefore, MALAT1 (together with SETD2) might act as scaffold to facility the interaction between pSmad2/3 and PPM1A, which lead to the dephosphorylation of pSmad2/3. This kinetically decrease of pSmad2/3 dephosphorylation might be the reason of increased Smads signaling activity in MALAT1 knockdown cells.
This work was supported by National Institutes of Health grants CA102325, CA106280, CA134568, DK077776 (to T.W.) and CA137729 (to C.H.)
Citation Format: Jinqiang Zhang, Chang Han, Tong Wu. The inhibitory effect of long non-coding RNA MALAT1 on Smads signaling pathway. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1825. doi:10.1158/1538-7445.AM2013-1825
Abstract 1827: HnRNP I interacts with lncRNA UCA1 and regulates its stability.
Authors: Huang Jianguo , Mo Yin-Yuan 
Published: August 2014

Abstract
Background: The human genome also makes a large number of non-coding RNAs including long non-coding RNAs (lncRNAs) and microRNAs in addition to protein-coding genes. While microRNAs are well characterized, little is known about lncRNAs. Emerging evidence has implicated lncRNAs in gene regulation; and they are often aberrantly expressed in a variety of human diseases, in particular cancer. The heterogeneous nuclear ribonucleoprotein I (hnRNP I) is a member of the large hnRNP family and has been implicated in mRNA splicing, stability and translational regulation of certain RNA transcripts. During screening for lncRNA-interacted proteins, we identified hnRNP I as a binding partner for UCA1. However, it remains unclear how hnRNP I interacts with UCA1 and whether such an interaction impacts UCA1-mediated cell growth and responsiveness to anti-cancer drugs.
Methods: To determine cell growth and response to doxorubicin, we performed MTT assays as well as cell counting assays. Protein levels from treated cells were analyzed by western blot. Real-time RT-PCR was used to determine relative expression of genes. Interaction between hnRNP I and UCA1 was confirmed by RNA co-immunoprecipitation and RNA pulldown. RNA stability was determined by treating MCF-7 cells with actinomycin D, followed by real-time RT-PCR. hnRNP I was knock down by transient transfection of hnRNP I siRNAs.
Results: We found that UCA1 induced proliferation and increased resistance to doxorubicin in MCF-7 cells. RNA pulldown assays revealed that UCA1 RNA directly interacted with hnRNP I. This interaction was further confirmed by RNA immunoprecipitation by hnRNP I antibody. Real-time RT-PCR revealed that knockdown of hnRNP I by RNAi reduced the level of UCA1 in MCF-7 cells and it also repressed the induction of UCA1 by doxorubicin. Further experiments suggested that hnRNP I impacts the UCA1 stability. We showed that half-life of UCA1 was about 12 h but was decreased to about 6 h in the presence of hnRNP I siRNAs. We further showed that the phosphorylated form of hnRNA I, predominantly in the cytoplasm, was a major form that interacts with UCA1.
Conclusion: Taken together, these results suggest that hnRNP I plays a role in regulation of the stability of UCA1. Since previous work implicates the involvement of UCA1 in tumor cell growth and chemoresistance, a better understanding of the underlying mechanism will provide new insight of targeting UCA1 in cancer therapy.
Citation Format: Jianguo Huang, Yin-Yuan Mo. HnRNP I interacts with lncRNA UCA1 and regulates its stability. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1827. doi:10.1158/1538-7445.AM2013-1827
Abstract 1831: Functional analysis of a novel long ncRNA involved in the tumorigenicity of colorectal cancer.
Authors: Taniue Kenzui , Kurimoto Akiko , Sugimasa Hironobu , Nasu Emiko , Nagashima Takeshi , Okada-Hatakeyama Mariko , Akiyama Tetsu 
Published: August 2014

Abstract
In recent years, the advent of high-throughput sequencing technologies has revealed that in addition to protein-coding genes, there are an even larger number of RNA transcripts that do not encode proteins and are instead defined as noncoding RNAs (ncRNAs). Numerous ncRNAs are misregulated in various diseases, especially cancer. We therefore attempted to identify and characterize novel long ncRNAs that regulate the tumorigenicity of colorectal cancer. Here we show that a novel long ncRNA, termed NR-X, is required for proliferation and tumorigenicity of colorectal cancer cells. We further show that NR-X is upregulated under hypoxic conditions and induces expression of hypoxia-responsive genes. Moreover, we succeeded to identify a chromatin-related protein that is associated with NR-X. In this presentation, we would like to present our data on the molecular functions of NR-X and discuss its significance in tumorigenesis.
Citation Format: Kenzui Taniue, Akiko Kurimoto, Hironobu Sugimasa, Emiko Nasu, Takeshi Nagashima, Mariko Okada-Hatakeyama, Tetsu Akiyama. Functional analysis of a novel long ncRNA involved in the tumorigenicity of colorectal cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1831. doi:10.1158/1538-7445.AM2013-1831
Abstract 1837: Exosomal non-coding RNAs as mediators for cell-cell communications .
Authors: Singh Ramesh , Mo Yin-Yuan 
Published: August 2014

Abstract
Introduction: Exosomes are 30-100nm membrane vesicles of endocytic origin containing proteins, mRNAs, microRNAs and lincRNAs. MicroRNAs are small and lincRNAs are large intergenic non-coding RNAs. They are emerging as key regulators of diverse cellular processes including cancer. In this study, we investigated the exosome-mediated transfer of microRNAs and lincRNAs, regulation and functional significance in breast cancer.
Approach: Exosomes were isolated by ultracentrifugation and stained with PKH26 dye for cellular uptake in different cells by confocal microscopy. This uptake was also shown by exosome isolated from stable 293T cell line overexpressing the pCDH-CD63-GFP fusion protein. To determine the effect of nSMase2 on exosome secretion, stable cell lines were established with smpd3 gene. Levels of microRNAs and lincRNAs secreted in media were determined by real time PCR and their function validated through western blot and cell invasion.
Results: We first showed that exosomes can be transferred among different cell lines through direct uptake. We found that miR-10b is highly expressed in metastatic breast cancer MDA-MB-231 cells as compared to non-metastatic breast cells. Also, nSMase2 promoted the exosome-mediated miR-10b secretion whereas ceramide inhibitor suppressed this secretion. Importantly, western blot revealed that upon uptake, miR-10b can still reduce the protein level of its target genes. We also found lincRNA 21A and BC200 to be secreted through exosome in cell type dependent manner with functional significance.
Conclusion: We found that the exosomes secreted from cancer cells containing microRNA and lincRNA can be taken up by other cells and have functional role in the recipient cells suggesting the importance of exosome-mediated RNA transfer in tumor microenvironment.
Citation Format: Ramesh Singh, Yin-Yuan Mo. Exosomal non-coding RNAs as mediators for cell-cell communications . [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1837. doi:10.1158/1538-7445.AM2013-1837
Abstract 1839: Long non-coding RNA SS-001 regulates proliferation, cell cycle, and migration in esophageal adenocarcinoma cells.
Authors: Song Jee Hoon , Meltzer Stephen J. 
Published: August 2014

Abstract
Esophageal adenocarcinoma (EAC) is one of the most rapidly increasing cancers in Western countries, but its underlying molecular mechanisms have not yet been completely elucidated. Recently, the discovery of long non-coding RNAs (lncRNAs) has added a new layer of complexity to the molecular architecture, identifying these molecules as emerging key regulators of diverse biological pathways. Increasing evidence shows that lncRNA dysregulation can lead to diseases such as cancer, but their potential involvement in EAC is not yet well-understood.
In this study, we identified a set of lncRNAs that were differentially expressed in esophageal cancers vs. normal tissues by massively parallel RNA sequencing. Expression levels were validated using quantitative real-time PCR. One lncRNA, designated SS-001, was significantly overexpressed and hypomethylated in EAC cell lines and tissues vs. normal esophageal epithelial cells and adjacent non-tumor tissues, respectively. SS-001 is an intergenic lncRNA located on chromosome 21.
In vitro assays in EAC cell lines demonstrated that knockdown of SS-001 resulted in inhibition of cell proliferation, the cell cycle, and migration. Moreover, patients with high SS-001 expression levels had a relatively poor prognosis. Our findings suggest that SS-001 may play a significant role in the development or progression of EAC and has the potential to serve as a prognostic biomarker.
Citation Format: Jee Hoon Song, Stephen J. Meltzer. Long non-coding RNA SS-001 regulates proliferation, cell cycle, and migration in esophageal adenocarcinoma cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1839. doi:10.1158/1538-7445.AM2013-1839
Abstract 1857: Dichloroacetate (DCA), pyruvate dehydrogenase kinase (PDK) inhibitor, enhances the cytotoxic effect induced by metformin .
Authors: Choi Yong Won , Lim In Kyoung 
Published: August 2014

Abstract
It has been recently reported that metformin, widely used as an antidiabetic drug, reduces risk of cancer development in human and animals based on its inhibitory effects observed in the several cancer cells and mouse tumor models. Metformin shows anti-tumor activity indirectly by enhancing insulin sensitivity and decreasing the level of insulin and IGF in serum. And it was also reported that metformin has direct anti-tumor activity through AMPK activation and mTOR inhibition. We also observed that metformin induces cytotoxicity in several different types of cancer cell lines. However when metformin was treated in high glucose condition, cytotoxic effect of metformin was significantly reduced and AMPK activation and mTOR inhibition were not observed. Metformin increased glycolysis by inhibiting mitochondrial respiratory complex I activity weakly and also inhibiting pyruvate dehydrogenase activity more significantly. So cytotoxic effect induced by metformin was dependent on glucose availability. Significant mitochondrial ROS production was observed in metformin treated cancer cells. NADPHs are produced by enhanced glycolysis through the pentose phosphate pathway. Since reducing power of NADPH is important for protection from ROS induced damage, we hypothesized that enhanced glycolysis by inhibition of PDH activity observed in metformin treated cancer cells is a protective role from metformin induced ROS injury. So when PDH activity was enhance by dichloroacetate (DCA), pyruvate dehydrogenase kinase (PDK) inhibitor, ROS production was more increased and resulted in significant cell death.
Citation Format: Yong Won Choi, In Kyoung Lim. Dichloroacetate (DCA), pyruvate dehydrogenase kinase (PDK) inhibitor, enhances the cytotoxic effect induced by metformin . [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1857. doi:10.1158/1538-7445.AM2013-1857
Abstract 1858: Development of a multiplex panel of biomarkers to assess energy metabolism in cancer.
Authors: Balakrishnan Meenakshi P. , Parchment Ralph E. , Hollingshead Melinda G. , Tomaszewski Joseph E. , Kinders Robert J. , Doroshow James H. , Srivastava Apurva K. 
Published: August 2014

Abstract
Background: Cancer cells have an altered glucose metabolism, which not only permits them to survive under hypoxic conditions but confers a distinct growth advantage by sustaining higher proliferation rates, invasiveness, and subsequent distant metastasis. This unique difference in glucose metabolism from healthy cells opens up a selective mechanism by which to kill cancer cells, bypassing normal healthy cells. Several enzymes in glucose metabolism are attractive targets for development of first-in-class therapeutic agents. We have undertaken development of a multiplex panel involving biomarkers involved in glucose metabolism to support drug development and to interrogate glycolytic pathway in cancer patients. Methods: The panel includes quantitative determinations of protein levels of hexokinase-2 (HK2), pyruvate kinase isoform-2 (PKM2), lactate dehydrogenase alpha (LDHa), and phosphorylated pyruvate dehydrogenase E1-alpha (phospho-PDHE1a). The sandwich immunoassays were built on the Luminex® xMAP technology platform using MagPlex magnetic beads and cloned proteins are employed as calibrators. The panel is designed to measure protein concentrations in fractionated cell extracts from flash frozen tumor biopsies. Results: We have completed development of three sandwich immunoassays for HK2, PK-M2, and LDHa on Luminex platform. Suitable capture antibodies as well as phycoerythrin labeled detection antibodies have been identified and tested for sandwich formation and multiplexing using recombinant calibrators and cell lysates. The multiplex is currently standardized for optimal assay performance. Known pharmacological modulators (3-bromopyruvate, bortezumib, oxamate, dichloroacetate) of HK2, PKM2, LDHa, and PDHE1a were used in vitro to demonstrate expected directional changes in biomarkers in the panel. Conclusions: We have developed a multiplex panel to measure protein levels of three key molecules involved in energy metabolism in cancer cells. The multiplex panel provides a critical tool, in the clinical setting, to support ongoing therapeutic strategies aimed at targeting the energy provision pathway in cancer patients. Importantly, the panel can be used to survey metabolic phenotype of different tumors to identify type and stage of cancer most susceptible to inhibition of glycolysis. Funded by NCI Contract No HHSN261200800001E.
Citation Format: Meenakshi P. Balakrishnan, Ralph E. Parchment, Melinda G. Hollingshead, Joseph E. Tomaszewski, Robert J. Kinders, James H. Doroshow, Apurva K. Srivastava. Development of a multiplex panel of biomarkers to assess energy metabolism in cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1858. doi:10.1158/1538-7445.AM2013-1858
Abstract 1859: CYLD downregulation induces aerobic glycolysis in human oral squamous cell carcinoma cells.
Authors: Nakamura Takuya , Shinriki Satoru , Hayashi Mitsuhiro 
Published: August 2014

Abstract
Purpose: The hypermetabolic nature of cancer cells, especially their increased reliance on aerobic glycolysis which has been associated with more aggressive phenotype, is considered metabolic hallmarks of cancer cells including oral squamous cell carcinoma (OSCC) cells. However, its precise mechanisms remain unknown. Cylindromatosis (CYLD) is recognized as a tumor suppressor gene whereas little is available about its impact on cancer progression. Our unpublished data showed that lower CYLD expression was associated with poor prognosis in OSCC patients. The objective of our study was to address specific contributions of CYLD to the signature metabolic features of OSCC cells.
Materials and methods: We determined the level of glucose consumption as well as lactate production to evaluate the effects of CYLD knockdown by siRNA on aerobic glycolysis in human OSCC cell lines. In addition, we measured extracellular acidification and oxygen consumption rates in OSCC cells by using XF Extracellular Flux Analyzers.
Results: Downregulation of CYLD significantly suppressed cell proliferation through cell cycle arrest while it inhibited apoptosis induced by serum withdrawal in vitro. In addition, loss of CYLD led to the increase in both of glucose consumption and lactate production in several OSCC cell lines. Consistently, culture medium became more acidic by CYLD nockdown,compared with the control. Furthermore, glycolysis was promoted by CYLD downregulation whereas no significant difference was found in mitochondrial respiration.
Conclusions: Dysfunction of CYLD may contribute to aerobic glycolysis without apparent influence on mitochondrial respiration. Further investigation on the advantage of such increased glycolysis and the change in metabolome profiling may lead to development of effective therapeutic strategies for OSCC and other malignancies.
Citation Format: Takuya Nakamura, Satoru Shinriki, Mitsuhiro Hayashi. CYLD downregulation induces aerobic glycolysis in human oral squamous cell carcinoma cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1859. doi:10.1158/1538-7445.AM2013-1859
Abstract 1869: DCA and metformin synergistically induce apoptosis in breast cancer cells.
Authors: Haugrud Allison B. , Zhuang Yongxian , Miskimins Wilson K. 
Published: August 2014

Abstract
The unusual metabolism of aerobic glycolysis in cancer cells suggests that this phenomenon can be exploited for improved therapies. Metformin, a drug used to treat type 2 diabetics, has clinical evidence of cancer prevention, and some studies show that cancer patients have a better response when it is used with neoadjuvant therapy. One of the cellular targets of metformin is the inhibition of Complex I in the mitochondrial electron transport chain. This leads to an accumulation superoxide species that can cause oxidative damage to cells and eventually lead to cell death. Based on this, we hypothesized that drugs which promote mitochondrial metabolism will synergize with metformin to increase superoxide production and promote cell death of cancer cells. Cancer cells using aerobic glycolysis display a high level of glucose uptake that is processed through glycolysis to pyruvate, which is then converted to lactate and secreted from the cells. Diverting pyruvate to mitochondrial metabolism rather than its conversion to lactate was expected to enhance metformin cytotoxicity. Dichloroacetate (DCA) is well established drug that has been tested in humans for the treatment of certain metabolic diseases. DCA works through inhibition of pyruvate dehydrogenase kinase (PDK), thus reducing entry of pyruvate into mitochondrial oxidative phosphorylation pathways. Consequently DCA is expected to promote mitochondrial metabolism and reduce lactate secretion by cancer cells. We demonstrate that DCA and metformin synergistically kill cultured breast cancer cells although the drug combination has no effect on non-transformed MCF10A breast epithelial cells. The enhanced cell death of the breast cancer cells displayed is through apoptosis, as shown by caspase-dependent increases in cleaved PARP. DCA treatment was associated with reduced lactate secretion by the cells and increased pH of the growth medium which suggested it promoted oxidative phosphorylation. The combination of two drugs also promoted increased mitochondrial superoxide production compared to metformin alone which was associated with increased oxidative damage. These findings indicate that, as expected, DCA promotes metformin-mediated cell death by enhancing mitochondrial metabolism of pyruvate. Therefore, it is possible that the combination of metformin with DCA, or similar compounds, could enhance cancer therapy.
Citation Format: Allison B. Haugrud, Yongxian Zhuang, Wilson K. Miskimins. DCA and metformin synergistically induce apoptosis in breast cancer cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1869. doi:10.1158/1538-7445.AM2013-1869