Volume 4, Issue 12 (Winter 2018)                   Caspian.J.Neurol.Sci 2018, 4(12): 24-29 | Back to browse issues page

XML Print

Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:

Mashayekhi S, Saberi A, Salehi Z. Expression of Matrix Metalloproteinase-2 and -9 in Meningioma. Caspian.J.Neurol.Sci. 2018; 4 (12) :24-29
URL: http://cjns.gums.ac.ir/article-1-216-en.html
1- Neuroscience Research Center, Poursina Hospital, Guilan University of Medical Sciences, Rasht, Iran
2- PhD Department of Biology, Faculty of Sciences, University of Guilan, Rasht, Iran
Full-Text [PDF 984 kb]   (32 Downloads)     |   Abstract (HTML)  (146 Views)
Full-Text:   (32 Views)
Meningioma accounts for more than 30% of all intracranial tumors. The prevalence peaks between the age of 60 and 70 years, at a male: female ration of 2: 3. Nevertheless, the tumor may also occur in children [1]. According to the World Health Organization (WHO), meningioma is categorized into 3 grades. Criteria include cell type, mitotic activity, cellularity, necrosis, and brain invasion. WHO grade I or benign meningioma represent approximately 90% of all meningiomas. WHO grade II (atypical meningioma) represents approximately 5%-7% of all meningiomas. They show a mitotic index of ≥4 mitosis per 10 Higher-Power Fields (HPF). WHO grade III or anaplastic meningioma accounts for 3% of all meningiomas, show a mitotic index of ≥20 mitosis per 10 HPF [2]
A critical step in tumor progression and recurrence is the infiltrative invasion into the contiguous tissue. Tumor invasion and metastasis formation are major obstacles for successful cancer therapy. Metastasis is a complex multistep process that requires sequential interactions between the invasive cell and the Extracellular Matrix (ECM). The remodeling of the ECM is carried out by a family of enzymes known as Matrix Metalloproteinases (MMP). MMPs are the family of endopeptidases, present in normal healthy tissue and constitute a large group of multidomain, zinc dependent endopeptidases capable of hydrolyzing all protein components of the ECM [3]. They can collectively degrade any component of ECM and basement membrane, and their excessive activity has been linked to numerous pathologies mainly including, but not limited to, tumor invasion and metastasis [4]. The tumor microenvironment can be regulated by MMPs. It has been shown that increased expression of MMPs plays a crucial role in the development of several human malignancies [5]
MMPs are grouped into the following categories on the basis of their substrate specificity: 1. Interstitial collagenases (MMP-2 and MMP-9); and 2. Stromelysins (MMP-3, MMP-10 and MMP-11), which contain a unique transmembrane domain in their COOH terminal that localizes these MMPs on the cell surfaces [6]. Among them, MMP-2 and MMP-9 are expressed as pro-forms, activated by cleavage of their amino terminal, and are consequently involved in proteolysis of the ECM and basement membrane. The activation of MMP is regulated by gene expression, pro-enzyme activation and inhibition of active enzymes by their endogenous Tissue Inhibitors of Metalloproteinases (TIMPs). Specifically, TIMP1 inhibits MMP-9 activity while TIMP2 controls MMP-2 activity [7]. TIMPs are also involved in the regulation of angiogenesis in brain tumor progression [8, 9]. It was documented that MMPs play a pivotal role in the process of malignant progression and that inhibition of MMP-9 activity results in the reduction of tumor invasion and metastasis in animal models [10]. MMP-2 and MMP-9 were shown to play a direct role in angiogenesis which require for tumor growth and metastasis [11]. MMPs and TIMPs are extensively studied in the field of tumor invasion and metastasis. Of the MMP family, MMP-2 and MMP-9 also known as gelatinase A and gelatinase B, respectively, are frequently mentioned in brain tumor biology [12]
It was shown that biochemical brain modifications could be reflected in the biological fluids including serum, therefore, measurement of peptides and amino acids in the serum might identify biomarkers of meningioma and disease progression. Thus, it is important to analyze serum biochemistry to find a reliable biomarker. The aim of this study was to evaluate the concentration of MMP-2 and MMP 9 in the serum of patients with different grades of meningioma.
Materials and Methods
Patient samples

The study included the number of 66 normal control and 101 patients with different grades of meningioma (42 cases of grade I, 38 grade II and 21 grade III). The serum samples was recruited between March 2013 and August 2017 at the Departments of neurology and neurosurgery, in an academic hospital affiliated to Guilan University of Medical Sciences, in the north of Iran. Meningioma cases were confirmed by Magnetic Resonance Imaging (MRI) and histopathology according to the World Health Organization (WHO) classification [2]. Patients who had any history of cancer/intracranial surgery, and received either radiotherapy or chemotherapy before surgery were excluded. The physical examination and imaging results of the patients at presentation and follow-up were recorded. Clinical information was collected and regularly updated for the patients through follow-up and questionnaires. 
One hundred and thirty two healthy volunteers were randomly selected from annual check-up visitors at the same hospital during the similar time period. The controls with a self-reported history of central nervous system-related diseases or cancer and previously receiving radiotherapy/chemotherapy were excluded. The research protocol was performed in accordance with the Declaration of Helsinki 1964 (revised 2013) and approved by the Ethics Committee of the Guilan University of Medical Sciences for Approval of Research Involving Human Subjects. Informed consent was obtained from all study subjects after explanation of the nature and possible consequences of the study. Samples of serum from normal subjects (controls) and patients with meningioma were collected and stored at -70oC until used.
 Protein analysis: Total protein concentration and ELISA
The total concentration of proteins in serum and Cerebro-Spinal Fluid (CSF) was determined by the Bio-Rad protein assay based on the Bradford dye procedure. MMP-2 and MMP-9 in serum was measured using the sensitive two sided ELISA and antiserum against Human MMP2 ELISA Kit (ab100606), Human MMP9 ELISA Kit (ab100610), (Cambridge, UK) were first coated with 80 ng primary antibody per well in 0.1 M Tris buffer. After overnight incubation, the plates were blocked with EIA buffer (50 mM Tris, pH 7.5, 0.3 M NaCl, 0.1% Triton X-100, 1% BSA and 1% Gelatine). The samples and standards were placed in triplicate wells and incubated overnight at room temperature. After washing a biotinylated secondary antibody (8 ng/mL) was added per well and the incubation was carried out for 24 hours at room temperature. b-Galactosidase coupled to Avidin was then added and after 2 hours was followed by washing. Finally 200 lM 4-methylumbelliferyl-b-galactoside (Sigma, Poole, UK) in 50 mM sodium phosphate and 10 mM MgCl2 buffer were added and the amount of fluorescence was measured after 40 min incubation at 37oC using a fluorimeter (Dynatech).
Statistical analysis
All data presented are expressed as mean±Standard Error of the Mean (SEM). Statistical analysis was done using one-way ANOVA and only values with P≤0.05 were considered as significant (SPSS software Version: 24.0).
Total protein concentrations 

The total concentration of proteins in serum samples from patients with different grades of meningioma and normal subjects was measured by the Bio-Rad protein assay. Results obtained demonstrated that no significant changes in all serum samples, starting from grades I to III meningioma, was observed (0.36±0.02, 0.35±0.06, 0.36±0.06 g/l as compared to controls (0.35±0.06 g/l) (P=0.58, 0.87 and 0.71, respectively) (Figure 1). 
Analysis of MMP-2 and MMP-9 concentrations in the serum by ELISA 
Using ELISA, it was shown that the levels of MMP-2 and MMP-9 in the serum samples of patients with meningioma were higher than in normal controls. The results showed that all serum samples, presented MMP-2 and MMP-9 expression, whereas, starting from grades I to III meningioma, a significant increase of MMP-2 and MMP-9 protein expression was observed (MMP-2 serum levels of 296.27±54.78, 346.90±41.77, 384.18±2742 ng/mL as compared to controls (242.18±40.93) (P=0.01, 0.0001 and 0.00001, respectively) and MMP-9 serum levels of 62.81±18.93, 84.45±9.21, 103±19.74 ng/mL as compared to control group (44.54±8.45 ng/mL) (P=0.008, 0.000012 and 0.00017, respectively) (Figures 2 and 3). An increase concentration of MMP-2 and MMP-9 is correlated with advanced grades of meningioma (Table 1).

Meningioma is an extra-medullary, intradural tumor arising from meningiothelial arachnoid cap cells which form the outer layer of the arachnoid matter. Multiple genes have been shown to be associated with meningioma [12]. Several studies have reported deletion of chromosome 22q and of its associated gene NF2 in case of meningioma [13]. In addition to chromosomal alterations, changes to individual genes have also been observed in meningioma. It was shown that meningioma had up-regulated expression of MMP family of proteins involved in cell growth, angiogenesis and invasion [12]. In addition, transfection of small interfering RNA (siRNA) constructs to reduce or silence expression of MMP-9 gene reduces meningioma migration and invasion in vivo [14]. It has been suggested that the increased ration of MMP-9 to TIMP-1 might be associated with the meningioma progression [15]. MMPs might be associated with meningioma that have aggressive characteristics. They may play a role in the biological behavior, recurrence and prognosis of meningioma [16]. It was demonstrated that MMPs and their TIMPs play key role in the pathogenesis of meningioma [17]. Increased levels of MMP-2 and MMP-9 in grade 1 meningioma were shown as prognostic or predictive factors of recurrence [18]
MMPs play a central role in different physiological and pathological conditions [19-22]. Changes in the MMPs Cerebrospinal Fluid (CSF) and serum concentration were shown in the patients with meningitis, Creutzfeldt Jacob Disease (CJD), hydrocephalus and seizure [23-27]. It has been shown that MMPs and TIMPs play key role in cell proliferation, invasion and migration in different cancers including laryngeal carcinoma cells and ovarian cancer [28, 29]. It has been demonstrated that MMP-9 and MMP-2 may be correlated with invasiveness of pituitary adenomas [30]. It was also shown that a balance between MMPs and TIMPs has an important role to play in human brain tumors and their expression may be valuable markers for tumor malignancy and the imbalance between MMP-2 and TIMP-2 may have an important role in glioblastoma invasion by degrading the extracellular matrix [31, 32]
Changes in the MMP expression in many cancer tissues have been demonstrated [33, 34]. In this study we found that the levels of serum MMP-2 and MMP 9 levels were significantly increased in patients with meningioma. Moreover, among patients with meningioma, the concentration of MMP-2 and MMP 9 were significantly higher in patients with advanced grade of the disease.
Meningioma demonstrate more complex cytogenetic and molecular profile with activation of oncogenes, inactivation of tumor suppressor genes, and alterations in other genes involved in several molecular pathways [35]. Multidrug Resistance-associated Proteins (MRPs) were shown to be utilized in the treatment of aggressive meningioma [36]. Increased expression of MRP5 was reported in anaplastic but not in benign meningioma [37]. Deletions on chromosome 22 map to 22q12, has been seen in meningioma [38]. Another common tumor suppressor genes that have been associated with the development of meningioma is DAL-1 (differentially expressed in adenocarcinoma of the lung) [39]. Increased expression of IGF2 transcripts, along with decreased expression of IGFBP2, has been shown to be associated with meningioma development, and anaplastic grade [40].
It is concluded that MMP-2 and MMP 9 are a constant composition of human serum. The result of this study suggests that MMP-2, MMP 9, TIMP1 and TIMP2 serum concentrations may provide a reliable and practical indicator of malignant potential and tumor progression. It is also concluded that MMP-2 and MMP 9 might be involved in the pathophysiology of meningioma and the detection of serum MMP-2 and MMP 9 may be useful in classifying meningioma.
This study was supported by ethic committee of research in University of Guilan and Guilan University of Medical Sciences. This research did not receive any specific grant from funding agencies in the public, commercial or not-for-profit sectors.
Conflict of Interest
The authors have no conflicts of interest.

  1. Hortobágyi T, Bencze J, Varkoly G, Kouhsari MC, Klekner Á. Meningioma recurrence. Open Med. 2016; 11:168–73. doi: 10.1515/med-2016-0032 
  2. Louis DN, Perry A, Reifenberger G, von Deimling A, Figarella-Branger D, Cavenee WK, et al. The 2016 World Health Organization Classification of Tumors of the Central Nervous System: a summary. Acta Neuropathol. 2016; 131(6):803–20. doi: 10.1007/s00401-016-1545-1
  3. Thakur V, Bedogni B. The membrane tethered matrix metalloproteinase MT1-MMP at the forefront of melanoma cell invasion and metastasis. Pharmacol Res. 2016; 111:17–22. doi: 10.1016/j.phrs.2016.05.019 
  4. Mittal R, Patel AP, Debs LH, Nguyen D, Patel K, Grati M, et al. Intricate Functions of matrix metalloproteinases in physiological and pathological conditions. J Cell Physiol. 2016; 231(12):2599–621. doi: 10.1002/jcp.25430 
  5. Hida Y, Hamada J. Differential expressions of matrix metalloproteinases, a disintegrin and metalloproteinases, and a disintegrin and metalloproteinases with thrombospondin motifs and their endogenous inhibitors among histologic subtypes of lung cancers. Anticancer Agents Med Chem. 2012; 12(7):744–52. doi: 10.2174/187152012802650156
  6. Mignatti P, Rifkin DB. Plasminogen activators and matrixmetalloproteinases in angiogenesis. Enzym and Protein. 1996; 49(1-3):117–37. PMID: 8797002
  7. Brinckerhoff CE, Matrisian LM. Matrix metalloproteinases: A tail of a frog that became a prince. Nat Rev Mol Cell Biol. 2002; 3(3):207–14. doi: 10.1038/nrm763 
  8. Rojiani MV, Ghoshal-Gupta S, Kutiyanawalla A, Mathur S, Rojiani AM. TIMP-1 overexpression in lung carcinoma enhances tumor kinetics and angiogenesis in brain metastasis. J Neuropathol Exp Neurol. 2015; 74(4):293–304. doi: 10.1097/nen.0000000000000175 
  9. Kadoglou NP, Liapis CD. Matrix metalloproteinases: contribution to pathogenesis, diagnosis, surveillance and treatment of abdominal aortic aneurysms. Curr Med Res Opin. 2004; 20(4):419–32. doi: 10.1185/030079904125003143
  10. Lozonschi L, Sunamura M, Kobari M, Egawa S, Ding L, Matsuno S. Controlling tumor angiogenesis and metastasis of C26 murine colon adenocarcinoma by a new matrix metalloproteinase inhibitor, KB-R7785, in two tumor models. Cancer Res. 1999; 59(6):1252-8. PMID: 10096556
  11. Deryugina EI, Quigley JP. Tumor angiogenesis: MMP-mediated induction of intravasation- and metastasis-sustaining neovasculature. Matrix Biol. 2015; 44-46:94–112. doi: 10.1016/j.matbio.2015.04.004
  12. Siddique K, Yanamandra N, Gujrati M, Dinh D, Rao J, Olivero W. Expression of matrix metalloproteinases, their inhibitors, and urokinase plasminogen activator in human meningiomas. Int J Oncol. 2003; 22(2): 289-94. doi: 10.3892/ijo.22.2.289
  13. Tanaka N, Nishisho I, Yamamoto M, Miya A, Shin E, Karakawa K, et al. Loss of heterozygosity on the long arm of chromosome 22 in pheochromocytoma. Genes, Chromosomes and Cancer. 1992; 5(4):399–403. doi: 10.1002/gcc.2870050416
  14. Rao J, Olivero W, Gujrati M, Dinh D, Gondi C, Spomar D, et al. RNAi-mediated abrogation of cathepsin B and MMP-9 gene expression in a malignant meningioma cell line leads to decreased tumor growth, invasion and angiogenesis. Int J Oncol. 2007; 31(5):1039-50. doi: 10.3892/ijo.31.5.1039
  15. Paek S, Kim D, Park CK, Phi J, Kim Y, Im S, et al. The role of matrix metalloproteinases and tissue inhibitors of matrix metalloproteinase in microcystic meningiomas. Oncol Rep. 2006; 16(1):49-56. doi: 10.3892/or.16.1.49
  16. Karaarslan N, Gurbuz MS, Caliskan T, Ayan E, Aker FV, Berkman MZ. The effect of matrix metalloproteinase enzyme-3 on the prognosis and biological behaviour of meningiomas. Turk Neurosurg. 2016; 26(5):678-83. doi: 1019-5149.jtn.12807-14.1
  17. Coven İ, Ozer O, Ozen O, Şahin Fİ, Altinors N. Presence of matrix metalloproteinase–2 and tissue inhibitor matrix metalloproteinase–2 gene polymorphisms and immunohistochemical expressions in intracranial meningiomas. J Neurosurg. 2014; 121(6):1478–82. doi: 10.3171/2014.8.jns13515 
  18. Okuducu AF, Zils U, Michaelis SAM, Mawrin C, von Deimling A. Increased expression of avian erythroblastosis virus E26 oncogene homolog 1 in World Health Organization grade 1 meningiomas is associated with an elevated risk of recurrence and is correlated with the expression of its target genes matrix metalloproteinase-2 and MMP-9. Cancer. 2006; 107(6):1365–72. doi: 10.1002/cncr.22130
  19. Shapiro S, Miller A, Lahat N, Sobel E, Lerner A. Expression of matrix metalloproteinases, sICAM-1 and IL-8 in CSF from children with meningitis. J Neurol Sci. 2003; 206(1):43–8. doi: 10.1016/s0022-510x(02)00317-9
  20. Sulik A, Wojtkowska M, Oldak E. Elevated levels of MMP-9 and TIMP-1 in the cerebrospinal fluid of children with echovirus type 30 and mumps meningitis. Scand J Immunol. 2008; 68(3):323–7. doi: 10.1111/j.1365-3083.2008.02137.x
  21. Kettlun A, Collados L, García L, Cartier LA, Wolf ME, Mosnaim AD, et al. Matrix metalloproteinase profile in patients with Creuztfeldt-Jakob disease. Int J Clin Pract. 2003; 57(6):475-8. PMID: 12918885
  22. Killer M, Arthur A, Al-Schameri AR, Barr J, Elbert D, Ladurner G, et al. Cytokine and growth factor concentration in cerebrospinal fluid from patients with hydrocephalus following endovascular embolization of unruptured aneurysms in comparison with other types of hydrocephalus. Neurochem Res. 2010; 35(10):1652–8. doi: 10.1007/s11064-010-0226-z
  23. Li YJ, Wang ZH, Zhang B, Zhe X, Wang MJ, Shi ST, et al. Disruption of the blood–brain barrier after generalized tonic-clonic seizures correlates with cerebrospinal fluid MMP-9 levels. J Neuroinflammation. 2013; 10:80. doi: 10.1186/1742-2094-10-80 
  24. Toroghi F, Mashayekhi F, Montazeri V, Saeedi Saedi H. [Association between MMP9 promoter polymorphism and breast cancer progression in northwest of Iran (Persian)]. Arak Med Univ J. 2016; 19(3):46-53.
  25. Saberi A, Salehi Z, Naderinabi B, Hojaat Ansari S, Mashayekhi S. Genetic dimension of intervertebral disc degeneration: polymorphism of matrix metalloproteinase 1 and 3 in the north Iranian population. Turk Neurosurg. 2017. doi: 10.5137/1019-5149.jtn.19978-17.0
  26. Shabanipour S, Mashayekhi F, Bahadori MH, Soruri ZZ. The relationship between MMP-9 promoterpolymorphism and IVF outcome. Cell Mol Biol. 2015; 61(1):64-7. PMID: 25817348
  27. Toroghi F, Mashayekhi F, Montazeri V, Saeedi Saedi H, Salehi Z. Association of MMP-9 promoter polymorphism and breast cancer among Iranian patients. Eur J Oncol. 2017; 22(1):38-42.
  28. Chen XD, Tang SX, Zhang JH, Zhang LT, Wang YW. CIP2A, an oncoprotein, is associated with cell proliferation, invasion and migration in laryngeal carcinoma cells. Oncol Rep. 2017; 38(2):1005–12. doi: 10.3892/or.2017.5759
  29. Yi X, Guo J, Guo J, Sun S, Yang P, Wang J, et al. EZH2-mediated epigenetic silencing of TIMP2 promotes ovarian cancer migration and invasion. Sci Rep. 2017; 7(1): 3568. doi: 10.1038/s41598-017-03362-z 
  30. Liu HY, Gu WJ, Wang CZ, Ji XJ, Mu YM. Matrix metalloproteinase-9 and -2 and tissue inhibitor of matrix metalloproteinase-2 in invasive pituitary adenomas. Medicine. 2016; 95(24):e3904. doi: 10.1097/md.0000000000003904
  31. Kachra Z, Beaulieu E, Delbecchi L, Mousseau N, Berthelet F, Moumdjian R, et al. Expression of matrix metalloproteinases and their inhibitors in human brain tumors. Clin Exp Metastasis. 1999; 17(7):555–66. doi: 10.1023/a:1006760632766
  32. Pei J, Park IH, Ryu HH, Li SY, Li CH, Lim SH, et al. Sublethal dose of irradiation enhances invasion of malignant glioma cells through p53-MMP 2 pathway in U87MG mouse brain tumor model. Radiat Oncol. 2015; 10:164. doi: 10.1186/s13014-015-0475-8
  33. Alshenawy HA. Immunohistochemical expression of epidermal growth factor receptor, E-cadherin, and matrix metalloproteinase–9 in ovarian epithelial cancer and relation to patient deaths. Ann Diagn Pathol. 2010; 14(6):387–95. doi: 10.1016/j.anndiagpath.2010.05.005
  34. AbdElazeem MA, El-Sayed M. The pattern of CD44 and matrix metalloproteinase 9 expression is a useful predictor of ulcerative colitis–associated dysplasia and neoplasia. Ann Diagn Pathol. 2015; 19(6):369–74. doi: 10.1016/j.anndiagpath.2015.08.004
  35. Suvà ML, Louis DN. Next-generation molecular genetics of brain tumours. Curr Opin Neurol. 2013; 26(6):681–7. doi: 10.1097/wco.0000000000000027 
  36. Dallas S. Multidrug resistance-associated proteins: expression and function in the central nervous system. Pharmacol Rev. 2006; 58(2):140–61. doi: 10.1124/pr.58.2.3
  37. Alexiou GA, Goussia A, Ntoulia A, Zagorianakou P, Malamou-Mitsi V, Voulgaris S, et al. Immunohistochemical study of MRP5 expression in meningiomas. Cancer Chemother Pharmacol. 2012; 71(3):825–8. doi: 10.1007/s00280-012-2057-x 
  38. Trofatter J. A novel moesin-, ezrin-, radixin-like gene is a candidate for the neurofibromatosis 2 tumor suppressor. Cell. 1993; 72(5):791–800. doi: 10.1016/0092-8674(93)90406-g
  39. Nunes F, Shen Y, Niida Y, Beauchamp R, Stemmer-Rachamimov AO, Ramesh V, et al. Inactivation patterns of NF2 and DAL-1/4.1B (EPB41L3) in sporadic meningioma. Cancer Genet Cytogenet. 2005; 162(2):135–9. doi: 10.1016/j.cancergencyto.2005.04.003
  40. Wrobel G, Roerig P, Kokocinski F, Neben K, Hahn M, Reifenberger G, et al. Microarray-based gene expression profiling of benign, atypical and anaplastic meningiomas identifies novel genes associated with meningioma progression. Int J Cancer. 2005; 114(2):249–56. doi: 10.1002/ijc.20733
Type of Study: Research | Subject: Special
Received: 2018/02/24 | Accepted: 2018/02/24 | Published: 2018/02/24

Add your comments about this article : Your username or Email:
Write the security code in the box

Send email to the article author

© 2017 All Rights Reserved | Caspian Journal of Neurological Sciences

Designed & Developed by : Yektaweb