Downregulation of TES by Hypermethylation in Glioblastoma Reduces Cell Apoptosis and Predicts Poor Clinical Outcome

Overview

Journal Eur J Med Res

Publisher Biomed Central

Specialty General Medicine

Date 2014 Dec 16

PMID 25498217

Citations 7

Authors

Yu Bai

Quan-Geng Zhang

Xin-Hua Wang

Affiliations

Soon will be listed here.

Abstract

Background: Gliomas are the most common human brain tumors. Glioblastoma, also known as glioblastoma multiform (GBM), is the most aggressive, malignant, and lethal glioma. The investigation of prognostic and diagnostic molecular biomarkers in glioma patients to provide direction on clinical practice is urgent. Recent studies demonstrated that abnormal DNA methylation states play a key role in the pathogenesis of this kind of tumor. In this study, we want to identify a novel biomarker related to glioma initiation and find the role of the glioma-related gene.

Methods: We performed a methylation-specific microarray on the promoter region to identify methylation gene(s) that may affect outcome of GBM patients. Normal and GBM tissues were collected from Tiantan Hospital. Genomic DNA was extracted from these tissues and analyzed with a DNA promoter methylation microarray. Testis derived transcript (TES) protein expression was analyzed by immunohistochemistry in paraffin-embedded patient tissues. Western blotting was used to detect TES protein expression in the GBM cell line U251 with or without 5-aza-dC treatment. Cell apoptosis was evaluated by flow cytometry analysis using Annexin V/PI staining.

Results: We found that the TES promoter was hypermethylated in GBM compared to normal brain tissues under DNA promoter methylation microarray analysis. The GBM patients with TES hypermethylation had a short overall survival (P <0.05, log-rank test). Among GBM samples, reduced TES protein level was detected in 33 (89.2%) of 37 tumor tissues by immunohistochemical staining. Down regulation of TES was also correlated with worse patient outcome (P <0.05, log-rank test). Treatment on the GBM cell line U251 with 5-aza-dC can greatly increase TES expression, confirming the hypermethylation of TES promoter in GBM. Up-regulation of TES prompts U251 apoptosis significantly. This study demonstrated that both TES promoter hypermethylation and down-regulated protein expression significantly correlated with worse patient outcome. Treatment on the GBM cell line (U251) with 5-aza-dC can highly release TES expression resulting in significant apoptosis in these cells.

Conclusions: Our findings suggest that the TES gene is a novel tumor suppressor gene and might represent a valuable prognostic marker for glioblastoma, indicating a potential target for future GBM therapy.

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References

Ichimura K, Bolin M, Goike H, Schmidt E, Moshref A, Collins V . Deregulation of the p14ARF/MDM2/p53 pathway is a prerequisite for human astrocytic gliomas with G1-S transition control gene abnormalities. Cancer Res. 2000; 60(2):417-24. View

Moniz S, Martinho O, Pinto F, Sousa B, Loureiro C, Oliveira M . Loss of WNK2 expression by promoter gene methylation occurs in adult gliomas and triggers Rac1-mediated tumour cell invasiveness. Hum Mol Genet. 2012; 22(1):84-95. DOI: 10.1093/hmg/dds405. View

Tobias E, Hurlstone A, MacKenzie E, McFarlane R, Black D . The TES gene at 7q31.1 is methylated in tumours and encodes a novel growth-suppressing LIM domain protein. Oncogene. 2001; 20(22):2844-53. DOI: 10.1038/sj.onc.1204433. View

Coutts A, MacKenzie E, Griffith E, Black D . TES is a novel focal adhesion protein with a role in cell spreading. J Cell Sci. 2003; 116(Pt 5):897-906. DOI: 10.1242/jcs.00278. View

Griffith E, Coutts A, Black D . Characterisation of chicken TES and its role in cell spreading and motility. Cell Motil Cytoskeleton. 2004; 57(3):133-42. DOI: 10.1002/cm.10162. View

Chene L, Giroud C, Desgrandchamps F, Boccon-Gibod L, Cussenot O, Berthon P . Extensive analysis of the 7q31 region in human prostate tumors supports TES as the best candidate tumor suppressor gene. Int J Cancer. 2004; 111(5):798-804. DOI: 10.1002/ijc.20337. View

Chung R, Whaley J, Kley N, Anderson K, Louis D, Menon A . TP53 gene mutations and 17p deletions in human astrocytomas. Genes Chromosomes Cancer. 1991; 3(5):323-31. DOI: 10.1002/gcc.2870030502. View

Deimling A, Louis D, Schramm J, Wiestler O . Astrocytic gliomas: characterization on a molecular genetic basis. Recent Results Cancer Res. 1994; 135:33-42. DOI: 10.1007/978-3-642-85039-4_5. View

Henson J, Schnitker B, Correa K, Deimling A, Fassbender F, Xu H . The retinoblastoma gene is involved in malignant progression of astrocytomas. Ann Neurol. 1994; 36(5):714-21. DOI: 10.1002/ana.410360505. View

10.

Ueki K, Ono Y, Henson J, Efird J, Deimling A, Louis D . CDKN2/p16 or RB alterations occur in the majority of glioblastomas and are inversely correlated. Cancer Res. 1996; 56(1):150-3. View

11.

Ichimura K, Schmidt E, Goike H, Collins V . Human glioblastomas with no alterations of the CDKN2A (p16INK4A, MTS1) and CDK4 genes have frequent mutations of the retinoblastoma gene. Oncogene. 1996; 13(5):1065-72. View

12.

Li J, Yen C, Liaw D, Podsypanina K, Bose S, Wang S . PTEN, a putative protein tyrosine phosphatase gene mutated in human brain, breast, and prostate cancer. Science. 1997; 275(5308):1943-7. DOI: 10.1126/science.275.5308.1943. View

13.

Steck P, Pershouse M, Jasser S, Yung W, Lin H, Ligon A . Identification of a candidate tumour suppressor gene, MMAC1, at chromosome 10q23.3 that is mutated in multiple advanced cancers. Nat Genet. 1997; 15(4):356-62. DOI: 10.1038/ng0497-356. View

14.

Wang S, Puc J, Li J, Bruce J, Cairns P, Sidransky D . Somatic mutations of PTEN in glioblastoma multiforme. Cancer Res. 1997; 57(19):4183-6. View

15.

Sarti M, Sevignani C, Calin G, Aqeilan R, Shimizu M, Pentimalli F . Adenoviral transduction of TESTIN gene into breast and uterine cancer cell lines promotes apoptosis and tumor reduction in vivo. Clin Cancer Res. 2005; 11(2 Pt 1):806-13. View

16.

Drusco A, Zanesi N, Roldo C, Trapasso F, Farber J, Fong L . Knockout mice reveal a tumor suppressor function for Testin. Proc Natl Acad Sci U S A. 2005; 102(31):10947-51. PMC: 1182460. DOI: 10.1073/pnas.0504934102. View

17.

Mueller W, Nutt C, Ehrich M, Riemenschneider M, Deimling A, van den Boom D . Downregulation of RUNX3 and TES by hypermethylation in glioblastoma. Oncogene. 2006; 26(4):583-93. DOI: 10.1038/sj.onc.1209805. View

18.

Boeda B, Briggs D, Higgins T, Garvalov B, Fadden A, McDonald N . Tes, a specific Mena interacting partner, breaks the rules for EVH1 binding. Mol Cell. 2007; 28(6):1071-82. DOI: 10.1016/j.molcel.2007.10.033. View

19.

Gunduz E, Gunduz M, Beder L, Nagatsuka H, Fukushima K, Sutcu R . Downregulation of TESTIN and its association with cancer history and a tendency toward poor survival in head and neck squamous cell carcinoma. Arch Otolaryngol Head Neck Surg. 2009; 135(3):254-60. DOI: 10.1001/archoto.2008.560. View

20.

Kanu O, Mehta A, Di C, Lin N, Bortoff K, Bigner D . Glioblastoma multiforme: a review of therapeutic targets. Expert Opin Ther Targets. 2009; 13(6):701-18. DOI: 10.1517/14728220902942348. View