MiR-338-5p Promotes Glioma Cell Invasion by Regulating TSHZ3 and MMP2
- 352 Downloads
This study was designed to examine differential expression of miR-338-5p in gliomas and the role of miR-338-5p in glioma cell invasion via its potential target gene TSHZ3 encoding Teashirt zinc finger homobox 3, predicted by bioinformatics, and matrix metallopeptidase 2 (MMP2), the key pro-invasive protease overexpressed in gliomas. Quantitative real-time reverse transcription PCR (qRT-PCR) and Spearman correlation analysis were used to determine differential expressions of miR-338-5p and TSHZ3 in astrocytic gliomas of different grades (n = 35) and glioblastoma cell lines (U87 and U251) in comparison to non-neoplastic brain (NNB) tissues (n = 6). Western blotting was used to determine the protein levels of TSHZ3 and MMP2 in glioblastoma cell lines and Matrigel invasion assay to examine the role of miR-338-5p in cell invasiveness. The results showed that the expression of miR-338-5p, normalized to hsnRNA U6, was significantly higher in grade III and IV gliomas and glioblastoma cell lines compared to that in NNB and grade II gliomas, whereas TSHZ3 expression, normalized to GAPDH, was inversely related to miR-338-5p (R = −0.636, P < 0.01). Luciferase assays showed TSHZ3 to be a target gene of miR-338-5p. In both U87 and U251 cells, miR-338-5p mimics increased MMP2 and invasiveness of the cells. Overexpression of ectopic TSHZ3 suppressed the cell invasiveness and attenuated the pro-invasive effect of miR-338-5p mimics. Overall, our results showed that miR-338-5p has a function in promoting glioma cell invasion by targeting TSHZ3 suppression on MMP2. In conclusion, miR-338-5p is a possible potential biomarker for the diagnosis and target for therapy of high-grade glioma.
KeywordsGlioma MiR-338-5p TSHZ3 MMP2 Invasion
This study was supported by the National Natural Science Foundation of China (Grant No. NSFC81372689), the Foundation of the Health Department in Jiangsu Province (Grant No. K201106) and the Foundation of the Chinese Anti-cancer Association (CSNO-2013-MSD009).
- Caubit X, Thoby-Brisson M, Voituron N, Filippi P, Bevengut M, Faralli H, Zanella S, Fortin G, Hilaire G, Fasano L (2010) Teashirt 3 regulates development of neurons involved in both respiratory rhythm and airflow control. J Neurosci 30(28):9465–9476. doi: 10.1523/JNEUROSCI.1765-10.2010 CrossRefPubMedGoogle Scholar
- Du R, Petritsch C, Lu K, Liu P, Haller A, Ganss R, Song H, Vandenberg S, Bergers G (2008) Matrix metalloproteinase-2 regulates vascular patterning and growth affecting tumor cell survival and invasion in GBM. Neuro Oncol 10(3):254–264. doi: 10.1215/15228517-2008-001 CrossRefPubMedPubMedCentralGoogle Scholar
- Faralli H, Martin E, Core N, Liu QC, Filippi P, Dilworth FJ, Caubit X, Fasano L (2011) Teashirt-3, a novel regulator of muscle differentiation, associates with BRG1-associated factor 57 (BAF57) to inhibit myogenin gene expression. J Biol Chem 286(26):23498–23510. doi: 10.1074/jbc.M110.206003 CrossRefPubMedPubMedCentralGoogle Scholar
- Kondraganti S, Mohanam S, Chintala SK, Kin Y, Jasti SL, Nirmala C, Lakka SS, Adachi Y, Kyritsis AP, Ali-Osman F, Sawaya R, Fuller GN, Rao JS (2000) Selective suppression of matrix metalloproteinase-9 in human glioblastoma cells by antisense gene transfer impairs glioblastoma cell invasion. Cancer Res 60(24):6851–6855PubMedGoogle Scholar
- McBride DJ, Etemadmoghadam D, Cooke SL, Alsop K, George J, Butler A, Cho J, Galappaththige D, Greenman C, Howarth KD, Lau KW, Ng CK, Raine K, Teague J, Wedge DC, Cancer Study Group AO, Caubit X, Stratton MR, Brenton JD, Campbell PJ, Futreal PA, Bowtell DD (2012) Tandem duplication of chromosomal segments is common in ovarian and breast cancer genomes. J Pathol 227(4):446–455. doi: 10.1002/path.4042 CrossRefPubMedPubMedCentralGoogle Scholar
- Natrajan R, Mackay A, Wilkerson PM, Lambros MB, Wetterskog D, Arnedos M, Shiu KK, Geyer FC, Langerod A, Kreike B, Reyal F, Horlings HM, van de Vijver MJ, Palacios J, Weigelt B, Reis-Filho JS (2012) Functional characterization of the 19q12 amplicon in grade III breast cancers. Breast Cancer Res 14(2):R53. doi: 10.1186/bcr3154 CrossRefPubMedPubMedCentralGoogle Scholar
- Yang TQ, Lu XJ, Wu TF, Ding DD, Zhao ZH, Chen GL, Xie XS, Li B, Wei YX, Guo LC, Zhang Y, Huang YL, Zhou YX, Du ZW (2014) MicroRNA-16 inhibits glioma cell growth and invasion through suppression of BCL2 and the nuclear factor-kappaB1/MMP9 signaling pathway. Cancer Sci 105(3):265–271. doi: 10.1111/cas.12351 CrossRefPubMedPubMedCentralGoogle Scholar