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      TRIM8 downregulation in glioma affects cell proliferation and it is associated with patients survival

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          Abstract

          Background

          Human gliomas are a heterogeneous group of primary malignant brain tumors whose molecular pathogenesis is not yet solved. In this regard, a major research effort has been directed at identifying novel specific glioma-associated genes. Here, we investigated the effect of TRIM8 gene in glioma.

          Methods

          TRIM8 transcriptional level was profiled in our own glioma cases collection by qPCR and confirmed in the independent TCGA glioma cohort. The association between TRIM8 expression and Overall Survival and Progression-free Survival in TCGA cohort was determined by using uni-multivariable Cox regression analysis. The effect of TRIM8 on patient glioma cell proliferation was evaluated by performing MTT and clonogenic assays. The mechanisms causing the reduction of TRIM8 expression were explored by using qPCR and in vitro assays.

          Results

          We showed that TRIM8 expression correlates with unfavorable clinical outcome in glioma patients. We found that a restored TRIM8 expression induced a significant reduction of clonogenic potential in U87MG and patient’s glioblastoma cells. Finally we provide experimental evidences showing that miR-17 directly targets the 3′ UTR of TRIM8 and post-transcriptionally represses the expression of TRIM8.

          Conclusions

          Our study provides evidences that TRIM8 may participate in the carcinogenesis and progression of glioma and that the transcriptional repression of TRIM8 might have potential value for predicting poor prognosis in glioma patients.

          Electronic supplementary material

          The online version of this article (doi:10.1186/s12885-015-1449-9) contains supplementary material, which is available to authorized users.

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          Most cited references15

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          Genetic pathways to glioblastoma: a population-based study.

          We conducted a population-based study on glioblastomas in the Canton of Zurich, Switzerland (population, 1.16 million) to determine the frequency of major genetic alterations and their effect on patient survival. Between 1980 and 1994, 715 glioblastomas were diagnosed. The incidence rate per 100,000 population/year, adjusted to the World Standard Population, was 3.32 in males and 2.24 in females. Observed survival rates were 42.4% at 6 months, 17.7% at 1 year, and 3.3% at 2 years. For all of the age groups, younger patients survived significantly longer, ranging from a median of 8.8 months ( 80 years). Loss of heterozygosity (LOH) 10q was the most frequent genetic alteration (69%), followed by EGFR amplification (34%), TP53 mutations (31%), p16(INK4a) deletion (31%), and PTEN mutations (24%). LOH 10q occurred in association with any of the other genetic alterations and was predictive of shorter survival. Primary (de novo) glioblastomas prevailed (95%), whereas secondary glioblastomas that progressed from low-grade or anaplastic gliomas were rare (5%). Secondary glioblastomas were characterized by frequent LOH 10q (63%) and TP53 mutations (65%). Of the TP53 mutations in secondary glioblastomas, 57% were in hotspot codons 248 and 273, whereas in primary glioblastomas, mutations were more equally distributed. G:C-->A:T mutations at CpG sites were more frequent in secondary than primary glioblastomas (56% versus 30%; P = 0.0208). This suggests that the acquisition of TP53 mutations in these glioblastoma subtypes occurs through different mechanisms.
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            Identification and functional characterization of microRNAs involved in the malignant progression of gliomas.

            Diffuse astrocytoma of World Health Organization (WHO) grade II has an inherent tendency to spontaneously progress to anaplastic astrocytoma WHO grade III or secondary glioblastoma WHO grade IV. We explored the role of microRNAs (miRNAs) in glioma progression by investigating the expression profiles of 157 miRNAs in four patients with primary WHO grade II gliomas that spontaneously progressed to WHO grade IV secondary glioblastomas. Thereby, we identified 12 miRNAs (miR-9, miR-15a, miR-16, miR-17, miR-19a, miR-20a, miR-21, miR-25, miR-28, miR-130b, miR-140 and miR-210) showing increased expression, and two miRNAs (miR-184 and miR-328) showing reduced expression upon progression. Validation experiments on independent series of primary low-grade and secondary high-grade astrocytomas confirmed miR-17 and miR-184 as promising candidates, which were selected for functional analyses. These studies revealed miRNA-specific influences on the viability, proliferation, apoptosis and invasive growth properties of A172 and T98G glioma cells in vitro. Using mRNA and protein expression profiling, we identified distinct sets of transcripts and proteins that were differentially expressed after inhibition of miR-17 or overexpression of miR-184 in glioma cells. Taken together, our results support an important role of altered miRNA expression in gliomas, and suggest miR-17 and miR-184 as interesting candidates contributing to glioma progression.
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              TRIM8 modulates STAT3 activity through negative regulation of PIAS3.

              TRIM8 is a member of the protein family defined by the presence of a common domain structure composed of a tripartite motif: a RING-finger, one or two B-box domains and a coiled-coil motif. Here, we show that TRIM8 interacts with protein inhibitor of activated STAT3 (PIAS3), which inhibits IL-6-dependent activation of STAT3. Ectopic expression of TRIM8 cancels the negative effect of PIAS3 on STAT3, either by degradation of PIAS3 through the ubiquitin-proteasome pathway or exclusion of PIAS3 from the nucleus. Furthermore, expression of TRIM8 in NIH3T3 cells enhances Src-dependent tumorigenesis. These findings indicate that TRIM8 enhances the STAT3-dependent signal pathway by inhibiting the function of PIAS3.
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                Author and article information

                Contributors
                +39 0882 416350 , l.micale@operapadrepio.it
                c.fusco@operapadrepio.it
                a.fontana@operapadrepio.it
                r.barbano@operapadrepio.it
                bartolomeoaugello@gmail.com
                elena.denittis@libero.it
                m.copetti@operapadrepio.it
                mt.pellico@operapadrepio.it
                bmandriani@gmail.com
                dariococciadiferro@gmail.com
                pparrella@operapadrepio.it
                fazio@unicampus.it
                l.dimitri@operapadrepio.it
                v.dangelo@operapadrepio.it
                chiara.novielli@unimi.it
                lidia.larizza@unimi.it
                antonio.daga@istge.it
                +39 0882 416350 , g.merla@operapadrepio.it
                Journal
                BMC Cancer
                BMC Cancer
                BMC Cancer
                BioMed Central (London )
                1471-2407
                16 June 2015
                16 June 2015
                2015
                : 15
                : 470
                Affiliations
                [ ]Medical Genetics Unit, IRCCS Casa Sollievo della Sofferenza, Poliambulatorio Giovanni Paolo II, I-71013 San Giovanni Rotondo (FG), Italy
                [ ]Biostatistics Unit, IRCCS Casa Sollievo della Sofferenza, Poliambulatorio Giovanni Paolo II, I-71013 San Giovanni Rotondo (FG), Italy
                [ ]Laboratory of Oncology, IRCCS Casa Sollievo della Sofferenza, I-71013 San Giovanni Rotondo (FG), Italy
                [ ]Ph.D program in Experimental and Regenerative Medicine, University of Foggia, Foggia, Italy
                [ ]Pathology Unit, IRCCS Casa Sollievo della Sofferenza, I-71013 San Giovanni Rotondo (FG), Italy
                [ ]Neurosurgery Unit, IRCCS Casa Sollievo Della Sofferenza, San Giovanni Rotondo (FG), Italy
                [ ]Medical Genetics, Department of Health Sciences, Università degli Studi di Milano, Milan, Italy
                [ ]Laboratory of Medical Cytogenetics and Molecular Genetics, Istituto Auxologico Italiano, Milan, Italy
                [ ]Gene Transfer Lab; IRCSS Azienda Ospedaliera Universitaria San Martino-IST Istituto Nazionale per la Ricerca sul Cancro, Genoa, Italy
                Article
                1449
                10.1186/s12885-015-1449-9
                4468980
                26077989
                9d345e37-26ad-4b03-83c9-06ae4d3d979d
                © Micale et al. 2015

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 19 February 2015
                : 19 May 2015
                Categories
                Research Article
                Custom metadata
                © The Author(s) 2015

                Oncology & Radiotherapy
                trim8,glioblastoma,mir-17,cell proliferation
                Oncology & Radiotherapy
                trim8, glioblastoma, mir-17, cell proliferation

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