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      SIRT6 Depletion Suppresses Tumor Growth by Promoting Cellular Senescence Induced by DNA Damage in HCC

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          Abstract

          The role of Sirtuin 6 (SIRT6) as a tumor suppressor or oncogene in liver cancer remains controversial. Thus, we identified the specific role of SIRT6 in the progression of hepatocellular carcinoma (HCC). SIRT6 expression was significantly higher in HCC cell lines and HCC tissues from 138 patients than in an immortalized hepatocyte cell line, THLE-2 and non-tumor tissues, respectively. SIRT6 knockdown by shRNA suppressed the growth of HCC cells and inhibited HCC tumor growth in vivo. In addition, SIRT6 silencing significantly prevented the growth of HCC cell lines by inducing cellular senescence in the p16/Rb- and p53/p21-pathway independent manners. Microarray analysis revealed that the expression of genes involved in nucleosome assembly was apparently altered in SIRT6-depleted Hep3B cells. SIRT6 knockdown promoted G2/M phase arrest and downregulation of genes encoding histone variants associated with nucleosome assembly, which could be attributed to DNA damage. Taken together, our findings suggest that SIRT6 acts as a tumor promoter by preventing DNA damage and cellular senescence, indicating that SIRT6 represents a potential therapeutic target for the treatment of HCC.

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            SIRT6 links histone H3 lysine 9 deacetylation to NF-kappaB-dependent gene expression and organismal life span.

            Members of the sirtuin (SIRT) family of NAD-dependent deacetylases promote longevity in multiple organisms. Deficiency of mammalian SIRT6 leads to shortened life span and an aging-like phenotype in mice, but the underlying molecular mechanisms are unclear. Here we show that SIRT6 functions at chromatin to attenuate NF-kappaB signaling. SIRT6 interacts with the NF-kappaB RELA subunit and deacetylates histone H3 lysine 9 (H3K9) at NF-kappaB target gene promoters. In SIRT6-deficient cells, hyperacetylation of H3K9 at these target promoters is associated with increased RELA promoter occupancy and enhanced NF-kappaB-dependent modulation of gene expression, apoptosis, and cellular senescence. Computational genomics analyses revealed increased activity of NF-kappaB-driven gene expression programs in multiple Sirt6-deficient tissues in vivo. Moreover, haploinsufficiency of RelA rescues the early lethality and degenerative syndrome of Sirt6-deficient mice. We propose that SIRT6 attenuates NF-kappaB signaling via H3K9 deacetylation at chromatin, and hyperactive NF-kappaB signaling may contribute to premature and normal aging.
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              Senescence and apoptosis: dueling or complementary cell fates?

              In response to a variety of stresses, mammalian cells undergo a persistent proliferative arrest known as cellular senescence. Many senescence-inducing stressors are potentially oncogenic, strengthening the notion that senescence evolved alongside apoptosis to suppress tumorigenesis. In contrast to apoptosis, senescent cells are stably viable and have the potential to influence neighboring cells through secreted soluble factors, which are collectively known as the senescence-associated secretory phenotype (SASP). However, the SASP has been associated with structural and functional tissue and organ deterioration and may even have tumor-promoting effects, raising the interesting evolutionary question of why apoptosis failed to outcompete senescence as a superior cell fate option. Here, we discuss the advantages that the senescence program may have over apoptosis as a tumor protective mechanism, as well as non-neoplastic functions that may have contributed to its evolution. We also review emerging evidence for the idea that senescent cells are present transiently early in life and are largely beneficial for development, regeneration and homeostasis, and only in advanced age do senescent cells accumulate to an organism's detriment.
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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, CA USA )
                1932-6203
                8 November 2016
                2016
                : 11
                : 11
                : e0165835
                Affiliations
                [1 ]Department of Life Sciences, Pohang University of Science and Technology, Pohang, Gyeongbuk, Korea
                [2 ]Cbs Bioscience Inc., Daejeon, Korea
                [3 ]Donnelly Centre, Departments of Molecular Genetics and Computer Science, University of Toronto, Toronto, Ontario, Canada
                [4 ]Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada
                [5 ]Department of Surgery, Ajou University School of Medicine, Suwon, Korea
                [6 ]Division of Integrative Biosciences & Biotechnology, Pohang University of Science and Technology, Pohang, Gyeongbuk, Republic of Korea
                Institute of Pathology, GERMANY
                Author notes

                Competing Interests: This is an original paper, and none of the material has been published or submitted for publication in any other primary scientific journal. All authors are aware of the submission to PLoS One for publication, and have agreed on the final content. The authors declare no conflict of interest. JHK is employees of CbsBioscience, whose company provided support in the form of salary for her. There are no patents, products in development or marketed products to declare. This does not alter our adherence to all the PLOS ONE policies on sharing data and materials.

                • Conceptualization: NL JHK DKK HJW KYC KTK.

                • Data curation: DKK.

                • Formal analysis: NL JHK DKK HJW KYC KTK HGR.

                • Funding acquisition: KYC KTK NL.

                • Investigation: NL JHK DKK HJW KYC HGR SRK.

                • Methodology: NL JHK KYC.

                • Project administration: NL KYC.

                • Resources: JHK HJW KYC KTK HGR.

                • Supervision: NL KYC KTK.

                • Validation: NL KTK HGR SRK.

                • Visualization: NL JHK DKK HGR.

                • Writing – original draft: NL KYC HGR JHK.

                • Writing – review & editing: NL JHK DKK HJW KYC KTK.

                [¤a]

                Current address: Department of Molecular, Cell and Cancer Biology, University of Massachusetts Medical School, Worcester, Massachusetts, United States of America

                Article
                PONE-D-16-27902
                10.1371/journal.pone.0165835
                5100879
                27824900
                a708c121-0b2e-4d7c-b347-cfc6702bf502
                © 2016 Lee et al

                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
                : 28 July 2016
                : 18 October 2016
                Page count
                Figures: 6, Tables: 0, Pages: 19
                Funding
                Funded by: National Research Foundation of Korea (NRF) grant funded by the Korea government (MEST)
                Award ID: 2014R1A2A2A01002931
                Award Recipient :
                Funded by: Next-Generation BioGreen 21 Program, Rural Development Administration, Republic of Korea
                Award ID: PJ011216012016
                Award Recipient :
                This study was supported by a grant from the National Research Foundation of Korea (NRF) grant funded by the Korea government (MEST) (2014R1A2A2A01002931) and the Next-Generation BioGreen 21 Program, Rural Development Administration, Republic of Korea (PJ011216012016). The funders provided support in the form of salaries for authors [NL, KYC], but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. JHK is employees of CbsBioscience, whose company provided support in the form of salary for her, but did not have any additional role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Medicine and Health Sciences
                Oncology
                Cancers and Neoplasms
                Carcinomas
                Hepatocellular Carcinoma
                Medicine and Health Sciences
                Oncology
                Cancers and Neoplasms
                Gastrointestinal Tumors
                Hepatocellular Carcinoma
                Medicine and Health Sciences
                Gastroenterology and Hepatology
                Liver Diseases
                Hepatocellular Carcinoma
                Biology and life sciences
                Genetics
                DNA
                DNA damage
                Biology and life sciences
                Biochemistry
                Nucleic acids
                DNA
                DNA damage
                Research and Analysis Methods
                Database and Informatics Methods
                Biological Databases
                Mutation Databases
                Biology and Life Sciences
                Genetics
                Mutation
                Mutation Databases
                Biology and life sciences
                Biochemistry
                Proteins
                DNA-binding proteins
                Histones
                Biology and Life Sciences
                Cell Biology
                Cell Processes
                Cell Cycle and Cell Division
                Research and Analysis Methods
                Bioassays and Physiological Analysis
                Microarrays
                Research and Analysis Methods
                Model Organisms
                Animal Models
                Mouse Models
                Research and Analysis Methods
                Specimen Preparation and Treatment
                Staining
                Cell Staining
                Custom metadata
                All relevant data are within the paper and its Supporting Information files except for the microarray data. Microarray data were deposited in the GEO database ( http://www.ncbi.nlm.nih.gov/geo/, accession number: GSE75905).

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                Uncategorized

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