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      Long Non-Coding RNA H19 Promotes Glioma Cell Invasion by Deriving miR-675

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          Abstract

          H19 RNA has been characterized as an oncogenic long non-coding RNA (lncRNA) in breast and colon cancer. However, the role and function of lncRNA H19 in glioma development remain unclear. In this study, we identified that H19/miR-675 signaling was critical for glioma progression. By analyzing glioma gene expression data sets, we found increased H19 in high grade gliomas. H19 depletion via siRNA inhibited invasion in glioma cells. Further, we found H19 positively correlated with its derivate miR-675 expression and reduction of H19 inhibited miR-675 expression. Bioinformatics and luciferase reporter assays showed that miR-675 modulated Cadherin 13 expression by directly targeting the binding site within the 3′ UTR. Finally, introduction of miR-675 abrogated H19 knockdown-induced cell invasion inhibition in glioma cells. To our knowledge, it is first time to demonstrate that H19 regulates glioma development by deriving miR-675 and provide important clues for understanding the key roles of lncRNA-miRNA functional network in glioma.

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          Most cited references25

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          Long non-coding RNA ANRIL is required for the PRC2 recruitment to and silencing of p15(INK4B) tumor suppressor gene.

          A 42 kb region on human chromosome 9p21 encodes for three distinct tumor suppressors, p16(INK4A), p14(ARF) and p15(INK4B), and is altered in an estimated 30-40% of human tumors. The expression of the INK4A-ARF-INK4B gene cluster is silenced by polycomb during normal cell growth and is activated by oncogenic insults and during aging. How the polycomb is recruited to repress this gene cluster is unclear. Here, we show that expression of oncogenic Ras, which stimulates the expression of p15(INK4B) and p16(INK4A), but not p14(ARF), inhibits the expression of ANRIL (antisense non-coding RNA in the INK4 locus), a 3.8 kb-long non-coding RNA expressed in the opposite direction from INK4A-ARF-INK4B. We show that the p15(INK4B) locus is bound by SUZ12, a component of polycomb repression complex 2 (PRC2), and is H3K27-trimethylated. Notably, depletion of ANRIL disrupts the SUZ12 binding to the p15(INK4B) locus, increases the expression of p15(INK4B), but not p16(INK4A) or p14(ARF), and inhibits cellular proliferation. Finally, RNA immunoprecipitation demonstrates that ANRIL binds to SUZ12 in vivo. Collectively, these results suggest a model in which ANRIL binds to and recruits PRC2 to repress the expression of p15(INK4B) locus.
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            The H19 locus: role of an imprinted non-coding RNA in growth and development.

            The H19 gene produces a non-coding RNA, which is abundantly expressed during embryonic development and down-regulated after birth. Although this gene was discovered over 20 years ago, its function has remained unclear. Only recently a role was identified for the non-coding RNA and/or its microRNA partner, first as a tumour suppressor gene in mice, then as a trans-regulator of a group of co-expressed genes belonging to the imprinted gene network that is likely to control foetal and early postnatal growth in mice. The mechanisms underlying this transcriptional or post-transcriptional regulation remain to be discovered, perhaps by identifying the protein partners of the full-length H19 RNA or the targets of the microRNA. This first in vivo evidence of a functional role for the H19 locus provides new insights into how genomic imprinting helps to control embryonic growth.
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              The imprinted H19 noncoding RNA is a primary microRNA precursor.

              Although H19 was the first imprinted noncoding transcript to be identified, the function of this conserved RNA has remained unclear. Here, we identify a 23-nucleotide microRNA derived from H19 that is endogenously expressed in human keratinocytes and neonatal mice and overexpressed in cells transfected with human or mouse H19 expression plasmids. These data demonstrate that H19 can function as a primary microRNA precursor and suggest that H19 expression results in the post-transcriptional downregulation of specific mRNAs during vertebrate development.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2014
                23 January 2014
                : 9
                : 1
                : e86295
                Affiliations
                [1 ]Department of Neurosurgery, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China
                [2 ]Liver Transplantation Center, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China
                [3 ]Surgical Research Center, Medical School, Southeast University, Nanjing, China
                [4 ]Department of Neurosurgery, People's hospital of Xuancheng city, Anhui, China
                Beijing Tiantan Hospital, Capital Medical University, China
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Conceived and designed the experiments: YS JXZ. Performed the experiments: YS YYW WKL. Analyzed the data: WKL JQ PW. Contributed reagents/materials/analysis tools: NL PW. Wrote the paper: YS JXZ TT. Conceived of the study and participated in its design and coordination: YPY.

                Article
                PONE-D-13-42595
                10.1371/journal.pone.0086295
                3900504
                24466011
                d7ddeffe-ca1c-4b25-883d-e959959cfb84
                Copyright @ 2014

                This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 17 October 2013
                : 13 December 2013
                Page count
                Pages: 7
                Funding
                This work is supported by National High Technology Research and Development Program 863 (2012AA02A508), China National Natural Scientific Fund (81072078, 81172389 and 81101901), Jiangsu Province's Natural Science Foundation (BK2010580 and 2011847), Jiangsu Province's Key Discipline of Medicine (XK201117), Jiangsu Province's Medical Major Talent program (RC2011051), Program for Development of Innovative Research Team in the First Affiliated Hospital of NJMU and Provincial Initiative Program for Excellency Disciplines, Jiangsu Province. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology
                Biochemistry
                Nucleic Acids
                RNA
                Molecular Cell Biology
                Gene Expression
                Neuroscience
                Medicine
                Oncology
                Cancer Risk Factors
                Genetic Causes of Cancer
                Cancers and Neoplasms
                Neurological Tumors
                Glioma

                Uncategorized
                Uncategorized

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