Proteomics-based identification of differentially expressed genes in human gliomas: down-regulation of SIRT2 gene

doi:10.1016/j.bbrc.2003.08.029

Biochemical and Biophysical Research Communications

Volume 309, Issue 3, 26 September 2003, Pages 558-566

https://doi.org/10.1016/j.bbrc.2003.08.029 Get rights and content

Abstract

A number of chromosomal abnormalities including 19q deletions have been associated with the formation of human gliomas. In this study, we employed a proteomics-based approach to identify possible genes involved in glioma tumorigenesis which may serve as potential diagnostic molecular markers for this type of cancer. By comparing protein spots from gliomas and non-tumor tissues using two-dimensional (2D) gel electrophoresis, we identified 11 up-regulated proteins and four down-regulated proteins in gliomas. Interestingly, we also discovered that a group of cytoskeleton-related proteins are differentially regulated in gliomas, suggesting the involvement of cytoskeleton modulation in glioma pathogenesis. We then focused on the cytoskeleton-related protein, SIRT2 (sirtuin homologue 2) tubulin deacetylase, which was down-regulated in gliomas. SIRT2 is located at 19q13.2, a region known to be frequently deleted in human gliomas. Subsequent Northern blot analysis revealed that RNA expression of SIRT2 was dramatically diminished in 12 out of 17 gliomas and glioma cell lines, in agreement with proteomic data. Furthermore, ectopic expression of SIRT2 in glioma cell lines led to the perturbation of the microtubule network and caused a remarkable reduction in the number of stable clones expressing SIRT2 as compared to that of a control vector in colony formation assays. These results suggest that SIRT2 may act as a tumor suppressor gene in human gliomas possibly through the regulation of microtubule network and may serve as a novel molecular marker for gliomas. Additional proteins were also identified, whose function in gliomas was previously unsuspected.

Section snippets

Materials and methods

Brain tissues. Brain tumor tissues and non-tumor tissues were obtained from Tottori University Hospital with informed consent and approved by an Institutional Review Committee. Tissues were flash-frozen in liquid nitrogen immediately after surgical removal and stored at −80 °C. Diagnosis of glioma was confirmed histopathologically. Gliomas were then classified according to the WHO grading scheme for central nervous system tumors [8].

Cell culture, transfection of EGFP-SIRT2 expression plasmid,

2D gel separation of proteins

Glioma tissues derived from five individuals were classified according to the WHO grading system for central nervous system tumors. In this study, gliomas classified as grade II (G201), III (G301 and G302), and IV (G402 and G405) were processed for 2D electrophoresis to isolate candidate proteins that are differentially regulated in gliomas. We detected about 1000 protein spots on the 2D gels after SYPRO Ruby staining (Fig. 1). The 2D electrophoresis profiles and relative spot intensities

Discussion

In this study, we analyzed differentially expressed proteins in human gliomas using a proteomics-based approach. To our knowledge, this is the first report of the use of a proteomics-based approach for the study of human gliomas.

We identified 11 proteins that are overexpressed and four proteins that are down-regulated in this type of cancer. One of the down-regulated proteins was identified as SIRT2 tubulin deacetylase which is located on 19q13.2, a region whose frequent deletion in gliomas was

Acknowledgements

We thank M. Kameyama, H. Yamada, and I. Kishimoto for technical assistance. We also thank R. Nishigaki for computing assistance. This work was supported in part by grants from the Ministry of Health, Labour and Welfare and the Ministry of Education, Culture, Sports, Science and Technology of Japan. C.G.T.T. is supported by the Japanese Government (Monbukagakusho) Scholarship.

References (26)

D.A Vandeputte et al.
GOA, a novel gene encoding a ring finger B-box coiled-coil protein, is overexpressed in astrocytoma
Biochem. Biophys. Res. Commun.
(2001)
R Nishigaki et al.
An extra human chromosome 21 reduces mlc-2a expression in chimeric mice and Down syndrome
Biochem. Biophys. Res. Commun.
(2002)
E Formstecher et al.
PEA-15 mediates cytoplasmic sequestration of ERK MAP kinase
Dev. Cell
(2001)
M Asamoto et al.
Prohibitin gene is overexpressed but not mutated in rat bladder carcinomas and cell lines
Cancer Lett.
(1994)
B.J North et al.
The human Sir2 ortholog, SIRT2, is an NAD+-dependent tubulin deacetylase
Mol. Cell
(2003)
D.W Cleveland et al.
Centromeres and kinetochores: from epigenetics to mitotic checkpoint signaling
Cell
(2003)
Y Gu et al.
Evidence that collapsin response mediator protein-2 is involved in the dynamics of microtubules
J. Biol. Chem.
(2000)
D Mornet et al.
Neurocalcin–actin interaction
Biochim. Biophys. Acta
(2001)
C.G Vedoy et al.
Isolation and characterization of genes associated with the anti-tumor activity of glucocorticoids
Brain Res. Mol. Brain Res.
(2002)
J Fueyo et al.
The functional role of tumor suppressor genes in gliomas: clues for future therapeutic strategies
Neurology
(1998)

B.K Rasheed et al.

Molecular pathogenesis of malignant gliomas

Curr. Opin. Oncol.

(1999)

A von Deimling et al.

Deletion mapping of chromosome 19 in human gliomas

Int. J. Cancer

(1994)

E.M Michiels et al.

Genes differentially expressed in medulloblastoma and fetal brain

Physiol. Genomics

(1999)

Cited by (250)

Ligand-based design and synthesis of new trityl histamine and trityl cysteamine derivatives as SIRT2 inhibitors for cancer therapy
2024, European Journal of Medicinal Chemistry
The relentless pursuit of novel therapeutic agents against cancer has led to the identification of multiple molecular targets, among which Sirtuin 2 (SIRT2) has garnered significant attention. This study presents an extensive SAR study of our reported trityl scaffold-based SIRT2 inhibitors. This study encompasses a range of different medicinal chemistry approaches to improve the activity of the lead compounds TH-3 and STCY1. The rationally designed and synthesized structures were confirmed using NMR and high-resolution mass spectroscopy before performing SIRT2 inhibition assay, NCI60 cytotoxicity test, and cell cycle analysis. Indeed, our strategies afforded hitherto unreported SIRT2 inhibitors with high activity, particularly 2a, 4a, 7c, and 7f. Remarkably, the presence of a lipophilic para substitution on the phenyl group of a freely rotating or a locked trityl moiety enhanced activity SIRT2 inhibition. Concomitantly, the synthesized compounds showed prominent activity against different cancer lines from the NCI60 assay. Of interest, compound 7c stands out as a potent and highly selective antiproliferative agent against leukemia and colon cancer panels. Furthermore, 7c treatment resulted in cell cycle arrest in MCF-7 cells at G2 phase and did not cause in vitro DNA cleavage.
Selective inhibition of SIRT2: A disputable therapeutic approach in cancer therapy
2024, Bioorganic Chemistry
Sirtuin 2 (SIRT2) is involved in a wide range of processes, from transcription to metabolism to genome stability. Dysregulation of SIRT2 has been associated with the pathogenesis and progression of different diseases, such as cancer and neurodegenerative disorders. In this context, targeting SIRT2 activity by small molecule inhibitors is a promising therapeutic strategy for treating related conditions, particularly cancer. This review summarizes the regulatory roles and molecular mechanisms of SIRT2 in cancer and the attempts to evaluate potential antitumor activities of SIRT2-selective inhibitors by in vitro and in vivo testing, which are expected to deepen our understanding of the role of SIRT2 in tumorigenesis and progression and may offer important clues or inspiration ideas for developing SIRT2 inhibitors with excellent affinity and selectivity.
Sirtuins' control of autophagy and mitophagy in cancer
2021, Pharmacology and Therapeutics
Mammalian cells use a specialized and complex machinery for the removal of altered proteins or dysfunctional organelles. Such machinery is part of a mechanism called autophagy. Moreover, when autophagy is specifically employed for the removal of dysfunctional mitochondria, it is called mitophagy. Autophagy and mitophagy have important physiological implications and roles associated with cellular differentiation, resistance to stresses such as starvation, metabolic control and adaptation to the changing microenvironment. Unfortunately, transformed cancer cells often exploit autophagy and mitophagy for sustaining their metabolic reprogramming and growth to a point that autophagy and mitophagy are recognized as promising targets for ongoing and future antitumoral therapies. Sirtuins are NAD+ dependent deacylases with a fundamental role in sensing and modulating cellular response to external stresses such as nutrients availability and therefore involved in aging, oxidative stress control, inflammation, differentiation and cancer. It is clear, therefore, that autophagy, mitophagy and sirtuins share many common aspects to a point that, recently, sirtuins have been linked to the control of autophagy and mitophagy. In the context of cancer, such a control is obtained by modulating transcription of autophagy and mitophagy genes, by post translational modification of proteins belonging to the autophagy and mitophagy machinery, by controlling ROS production or major metabolic pathways such as Krebs cycle or glutamine metabolism. The present review details current knowledge on the role of sirtuins, autophagy and mitophagy in cancer to then proceed to discuss how sirtuins can control autophagy and mitophagy in cancer cells. Finally, we discuss sirtuins role in the context of tumor progression and metastasis indicating glutamine metabolism as an example of how a concerted activation and/or inhibition of sirtuins in cancer cells can control autophagy and mitophagy by impinging on the metabolism of this fundamental amino acid.
Sirtuins as NAD<sup>+</sup>-dependent deacetylases and their potential in medical therapy
2021, Medical Epigenetics
Deacetylases are a class of enzymes which remove the ɛ-N-acetyl groups from lysine amino acids in histone and nonhistone targets. There are four classes of HDACs, with a distinct subgroup called sirtuins (HDAC class III) that rely on nicotinamide adenine dinucleotide (NAD⁺) for their catalytic activity. The fact that most of the sirtuins require NAD⁺ for their deacetylase activity indicates that they are major energy-sensing proteins in cells and tissues with the potency to affect a plethora of biochemical, physiological, and pathological processes. These effects exerted by various sirtuins are discussed in this chapter, as well as the processes regulating NAD⁺ levels by de novo and salvage pathways. Finally, we elaborate on various agents that modulate salvage pathway enzymes and sirtuin activity, and their preclinical and clinical implications in health and disease.
Sirtuin signaling in hematologic malignancies
2021, Sirtuin Biology in Cancer and Metabolic Disease: Cellular Pathways for Clinical Discovery
The sirtuins are a class of NAD⁺-dependent enzymes that are critical mediators of health maintenance and aging. Interestingly, sirtuins regulate many processes that are hijacked by cancer cells, including genomic stability and metabolism. This chapter summarizes each of the seven human sirtuins and their functions, particularly as it relates to signaling in hematologic malignancies. The chapter ends by discussing therapeutic strategies to exploit sirtuin signaling to better treat hematologic malignancies. Throughout, the chapter highlights interesting unanswered questions involving sirtuin biology in hematologic malignancies.
Sirtuin Inhibitor Cambinol Induces Cell Differentiation and Differently Interferes with SIRT1 and 2 at the Substrate Binding Site
2023, Biomedicines

View all citing articles on Scopus

View full text

Proteomics-based identification of differentially expressed genes in human gliomas: down-regulation of SIRT2 gene

Abstract

Section snippets

Materials and methods

2D gel separation of proteins

Discussion

Acknowledgements

Biochem. Biophys. Res. Commun.

Biochem. Biophys. Res. Commun.

Dev. Cell

Cancer Lett.

Mol. Cell

Cell

J. Biol. Chem.

Biochim. Biophys. Acta

Brain Res. Mol. Brain Res.

The functional role of tumor suppressor genes in gliomas: clues for future therapeutic strategies