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|Year : 2014 | Volume
| Issue : 1 | Page : 37-41
Assessment of expression of epidermal growth factor receptor and p53 in meningiomas
Swetha Narla1, Megha S Uppin1, M Vijaya Saradhi2, BP Sahu2, AK Purohit2, C Sundaram1
1 Department of Pathology, Nizam's Institute of Medical Sciences, Hyderabad, Andhra Pradesh, India
2 Department of Neurosurgery, Nizam's Institute of Medical Sciences, Hyderabad, Andhra Pradesh, India
|Date of Submission||13-May-2013|
|Date of Decision||23-Dec-2013|
|Date of Acceptance||26-Jan-2014|
|Date of Web Publication||7-Mar-2014|
Department of Pathology, Nizam's institute of Medical Sciences, Hyderabad - 500 082, Andhra Pradesh
Source of Support: None, Conflict of Interest: None
Background: Meningiomas represent about 30% of primary adult central nervous system tumors. Though slow growing, they recur, causing significant morbidity and mortality. Objective: The objective of the following study is to grade meningiomas according to World Health Organization (WHO) 2007 criteria and to correlate the grade with degree of expression of epidermal growth factor receptor (EGFR) and p53. Materials and Methods: Meningiomas diagnosed in the year 2010 in the Department of Pathology of our institute, were included in the study. Clinical and radiological findings were noted from medical records. The histopathology slides were reviewed and the tumors were graded according to WHO 2007 criteria. Tissue microarrays (TMA) were prepared and immunohistochemical analysis with epithelial membrane antigen, Vimentin, Ki67, EGFR and p53 was performed on the TMA slides. Results: A total of 79 meningiomas diagnosed during the study period included 30 male and 49 female patients with an age range of 10-75 years. There was a female preponderance with M:F ratio of 1:1.63. EGFR was found to be higher in grade I (82.93%) compared with grade II (35.71%) and grade III tumors (20%) with an overall positivity of 60.81%. Mean p53 indices were higher in grade III (50%) compared with grade II (39.29%) and grade I tumors (38.46%) with an overall positivity of 39.44%. Ki67 labeling index (LI) was significantly high in grade III (16.4%) compared with grade II (6.46%) and grade I tumors (3.13%). Conclusion: EGFR expression and Ki67 LI correlated with grade of meningioma P < 0.0001 and P < 0.0001 respectively which were statistically significant whereas p53 expression did not correlate (P - 0.90).
Keywords: Epidermal growth factor receptor, grade, Ki67 labeling index, meningioma, p53, tissue microarrays
|How to cite this article:|
Narla S, Uppin MS, Saradhi M V, Sahu B P, Purohit A K, Sundaram C. Assessment of expression of epidermal growth factor receptor and p53 in meningiomas. Neurol India 2014;62:37-41
|How to cite this URL:|
Narla S, Uppin MS, Saradhi M V, Sahu B P, Purohit A K, Sundaram C. Assessment of expression of epidermal growth factor receptor and p53 in meningiomas. Neurol India [serial online] 2014 [cited 2020 Feb 22];62:37-41. Available from: http://www.neurologyindia.com/text.asp?2014/62/1/37/128276
| » Introduction|| |
Meningiomas represent about 30% of primary adult central nervous system tumors. They are generally slow growing tumors, but can recur and cause significant morbidity and mortality.  Primary therapy for meningiomas is surgery. Overall recurrence rate has been reported to be around 20%. , The high recurrence rate in partially resected meningiomas has given way to the use of additional therapy. There are ongoing efforts to develop new treatment modalities for meningiomas based on the genetic pathways and receptors. One such marker is epidermal growth factor receptor (EGFR) which has been described in breast, lung, head and neck, glioblastoma and colorectal cancers. , Weisman, et al.  characterized expression of EGFR in meningiomas and suggested that EGFR is involved in the proliferation and/or differentiation of meningothelial cells. Although, there are a few studies from the west ,, on the expression of EGFR assessment in meningiomas, until date there are no studies from India. Another such marker is p53 which is a tumor suppressor gene located on the short arm of chromosome 17. The function of p53 is critical to the efficiency of many cancer treatment procedures. Consequently, tumors which carry mutations in p53 are often difficult to treat and their prognosis is poor. Studies of p53 expression in meningiomas are limited. ,,, This study was designed to determine whether meningiomas express EGFR and p53 and if so, to establish a correlation between the histopathologic grade of these tumors and the degree of expression.
| » Materials and Methods|| |
This was a retrospective study. All cases of meningiomas diagnosed in the year 2010 in the department of pathology were included in the study. All hematoxylin and eosin stained slides were reviewed and graded according to World Health Organization (WHO) criteria 2007.  Presence or absence of brain invasion was noted. The best section which was free from necrosis or hemorrhage was selected for tissue microarray (TMA). Representative areas from each section were marked on the tissue block from which 3 cores were extracted. Two TMA blocks were prepared from the paraffin blocks by manual method. One TMA block consisted of 120 cores from 40 blocks and the other TMA block consisted of 117 cores from 39 blocks.
Immunohistochemical analysis with epithelial membrane antigen (EMA), Vimentin, Ki67 (ready to use Biogenex, USA), EGFR (monoclonal mouse antibody, 1:100, Biogenex, USA) and p53 (monoclonal antibody, 1:50, DAKO, USA) was performed by horseradish peroxidase polymer technique on the TMA slides. Cores were recorded as non-informative if the tissue was lost in processing/there was no recognizable tumor in the core/or there were extensive staining artefacts (e.g. inappropriate staining of collagen or tissue edges and tissue creases in a specimen with minimal tissue retained). EGFR expression was considered positive if it stained either cytoplasm/cell membrane and was scored according to the percentage of stained cells.
Scoring for p53 nuclear staining was based on a four-point scale: no staining 0; <5% of nuclei with positive staining 1; 5-30% of nuclei with positive staining 2; and 30% of nuclei with positive staining 3. For the purpose of analysis, all p53 scores of 0 and 1 were considered as "negative," whereas the scores of 2 and 3 were considered as "positive." For counting Ki67 labeling index (LI) and p53, 100 nuclei/core were counted and an average was taken and for counting EGFR, 100 cells/core were counted and an average was taken. Comparisons of immunoreactivity frequencies of EGFR and p53 between tumor grades were carried out using the Chi-square test with Yates' correction.
| » Results|| |
A total of 100 cases of meningiomas were diagnosed in our department during the study period. Only 79 cases were included in the present study where sufficient material was available in the blocks. These included 30 male and 49 female patients with M:F ratio of 1:1.63. The age ranged from 10 years to 75 years (mean 46.27 years). There were 46 grade I meningiomas [Figure 1]a (58.23%), 28 grade II meningiomas [Figure 2]a (35.44%) and 5 grade III meningiomas [Figure 3]a (6.33%). There were 70 (88.61%) intracranial and 9 (11.39%) spinal meningiomas. Among 79 cases of meningiomas, transitional meningiomas were the most common subtype both in intracranial (31/70; 44.29%) and spinal (9/9; 100%) location. [Table 1] shows the demographic data, clinical features and IHC results of meningiomas.
|Figure 1: Grade I meningioma (a) grade I (transitional) meningioma (H and E, ×100). (b) Tumor cells stained positive with epidermal growth factor receptor (×100). (c) Nuclear staining with p53 (×100). (d) Nuclear staining with Ki67 (×100)|
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|Figure 2: Grade II meningioma (a) grade II meningioma-high cellularity with loss of architectural pattern, mitosis (H and E, ×100). (b) Tumor cells stained positive with epidermal growth factor receptor (×100). (c) Nuclear staining with p53 (×100). (d) Nuclear staining with Ki67 (×100)|
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|Figure 3: Grade III meningioma (a) grade III (rhabdoid) meningioma (H and E, ×100). (b) Tumor cells stained positive with epidermal growth factor receptor (×100). (c) Nuclear staining with p53 (×100). (d) Nuclear staining with Ki67 (×100)|
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|Table 1: Summary of demographic data, clinical features and immunohistochemical results of meningiomas|
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The final analysis for IHC was carried out on EMA (79 cases), Vimentin (79 cases), EGFR (74 cases), p53 (71 cases) and Ki67 LI (79 cases) as some of the cores showed edge artefacts and some were lost in processing.
IHC with EMA and Vimentin
EMA and Vimentin immunopositivity was seen in all the cases and in all grades of meningiomas (79/79; 100% positivity).
IHC with EGFR
EGFR expression was higher in grade I tumors (82.93%) [Figure 1]b when compared with grade II (35.71%) [Figure 2]b and grade III tumors (20%) [Figure 3]b with an overall positivity of 60.81% respectively. These values showed statistical significance (P < 0.0001) on Chi-square analysis.
IHC with p53
Mean p53 indices were lower in grade I tumors (38.46%) [Figure 1]c compared with grade II (39.29%) [Figure 2]c and grade III meningiomas (50%) [Figure 3]c with an overall positivity of 39.44%. But these values did not show statistical significance (P = 0.90).
IHC with Ki67
The mean values of Ki67 LI increased from grade I (3.13%) [Figure 1]d to grade II (6.46%) [Figure 2]d to grade III tumors (16.4%) [Figure 3]d. These values were statistically significant (P ≤ 0.0001).
| » Discussion|| |
WHO has recognized three grades in meningiomas which helps to predict the clinical behavior of these tumors.  Grade I meningiomas have a low rate of recurrence with a long survival time. Grade II meningiomas are associated with a high rate of recurrence (40%). Grade III meningiomas are highly aggressive with a low survival time (<2 years). In our study there was a female preponderance among all grades of meningiomas similar to other studies.  Grade II and III meningiomas were also more common in females with M:F ratio of 1:2.5 whereas according to Bollag et al.,  grade II and grade III meningiomas were more common in men.
Despite many recent modifications in the histological classification of meningiomas, there is still considerable variability in the biological behavior within each of the tumor grades and hence further patient stratification is required. Expression of EGFR in meningiomas may provide useful prognostic information for patient stratification and treatment as it may be an opportunity for pharmacological intervention in the disease process. There have been a few studies which have studied the expression patterns of EGFR in various grades of meningiomas representing all the three grades of meningiomas as defined by the WHO 2007 criteria. ,,, In our study, EGFR expression was detected in 60.81% of all meningioma samples tested. It was found to be higher in grade I (82.93%) when compared with grade II (35.71%) and grade III meningiomas (20%). There was a significant association between percentages of EGFR staining and histopathologic grade (P = 0.0001). In the study by Wernicke et al.,  EGFR expression was noted in 86% of all meningiomas. Lusis et al.  showed that EGFR expression was seen in 88% of the incidentally detected meningiomas and only 20% of the grade I, 61% of grade II and 44% of grade III meningiomas demonstrated immunoreactivity for EGFR (P - 0.032). In other studies EGFR has been found in nearly 100% of meningioma specimens in tissue culture. ,,
Reports vary on EGFR expression in meningiomas, which range from 33% to 100% respectively [Table 2]. In majority of studies, the expression ranged between 50% and 60% respectively. ,,,, Different techniques were used in different studies, which may be the cause for this variation in expression rates. Andersson et al.  utilized immunohistochemical and quantitative real-time polymerase chain reaction to identify EGFR expression in 26 cases. They did not identify an association between expression level and outcome, but their study included only cases of grade I meningiomas. Manasa et al.  also carried out TMA IHC technique similar to our study but on the correlation of p53 and Ki67 expression with grade and subtype of ependymomas.
|Table 2: Summary of immunohistochemistry results with EGFR according to different studies|
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The present study was conducted with the objective to evaluate the expression of p53 and Ki67 in different grades of meningiomas. Chacko et al.  showed that grade II tumors had a higher MIB-1 LI than grade I tumors. The MIB-1 LI used in conjunction with histological features can help in making a recommendation regarding potentially aggressive behavior in meningiomas. Babu et al.  studied Ki67 LI in various histological subtypes and grades of meningioma and correlated it with various parameters for recurrence. They showed that high Ki67 LI correlated with higher grade of meningioma. Pratibha et al.  provided evidence of p53 alteration through allelic deletion that are common primary somatic mutation events which occur irrespective of grade and stage and are hence probably associated with an early phase of tumorigenesis and/or tumor progression in childhood sarcomas and retinoblastomas. Loss or mutations of wild type p53 have only a limited role in tumorigenesis of meningiomas.  Mean p53 indices were higher in grade III tumors (50%) compared with grade II (39.29%) and grade I meningiomas (38.46%) with an overall positivity of 39.44% in our study compared with 64% demonstrated by Perry et al.  Ohkoudo et al.  and other authors reported clinical significance for p53 expression in meningioma since an association was found between p53 immunoreactivity and histological type. ,,, Wang et al.  found p53 expression in 80% of grade II meningiomas, but in none of the grade I meningiomas investigated. Therefore they concluded that p53 could be considered as a marker for malignant transformation. However, other authors could not confirm these observations. ,
Aguiar et al.,  p53 expression was observed in 75% of grade II and grade III meningiomas and correlated with histological grade (P = 0.0009). [Table 3] shows a summary of previous studies of correlation of p53 with histological grade. The physiological significance of the role of EGFR in human meningioma is unknown at present, but knowledge of factors which regulate tumor growth can lead to a possible treatment when tumors are inoperable.  Application of EGFR targeted therapies to meningiomas requires further trials to be conducted. By detecting EGFR mutations, meningiomas could become eligible for targeted therapy. Immunohistochemical evidence of EGFR expression is a prognostic indicator in meningiomas.
|Table 3: Summary of previous studies of correlation of p53 with histological grade|
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| » Acknowledgments|| |
The authors gratefully acknowledge Dr. Sanjay Navani for helping in preparation of tissue microarray blocks.
| » References|| |
|1.||Louis DN, Scheithauer BW, Budka H, Deimling VA, Kepes JJ. Meningiomas. In: Kleihues P, Cavenee WK, editors. World Health Organization Classification of Tumors. Lyon: IARC; 2007. p. 164-72. |
|2.||Jääskeläinen J, Haltia M, Servo A. Atypical and anaplastic meningiomas: Radiology, surgery, radiotherapy, and outcome. Surg Neurol 1986;25:233-42. |
|3.||Mirimanoff RO, Dosoretz DE, Linggood RM, Ojemann RG, Martuza RL. Meningioma: Analysis of recurrence and progression following neurosurgical resection. J Neurosurg 1985;62:18-24. |
|4.||Salomon DS, Brandt R, Ciardiello F, Normanno N. Epidermal growth factor-related peptides and their receptors in human malignancies. Crit Rev Oncol Hematol 1995;19:183-232. |
|5.||Libermann TA, Razon N, Bartal AD, Yarden Y, Schlessinger J, Soreq H. Expression of epidermal growth factor receptors in human brain tumors. Cancer Res 1984;44:753-60. 5. |
|6.||Weisman AS, Raguet SS, Kelly PA. Characterization of the epidermal growth factor receptor in human meningioma. Cancer Res 1987;47:2172-6. |
|7.||Andersson U, Guo D, Malmer B, Bergenheim AT, Brännström T, Hedman H, et al. Epidermal growth factor receptor family (EGFR, ErbB2-4) in gliomas and meningiomas. Acta Neuropathol 2004;108:135-42. |
|8.||Carroll RS, Black PM, Zhang J, Kirsch M, Percec I, Lau N, et al. Expression and activation of epidermal growth factor receptors in meningiomas. J Neurosurg 1997;87:315-23. |
|9.||Johnson MD, Horiba M, Winnier AR, Arteaga CL. The epidermal growth factor receptor is associated with phospholipase C-gamma 1 in meningiomas. Hum Pathol 1994;25:146-53. |
|10.||Wang JL, Zhang ZJ, Hartman M, Smits A, Westermark B, Muhr C, et al. Detection of TP53 gene mutation in human meningiomas: A study using immunohistochemistry, polymerase chain reaction/single-strand conformation polymorphism and DNA sequencing techniques on paraffin-embedded samples. Int J Cancer 1995;64:223-8. |
|11.||Ellison DW, Lunec J, Gallagher PJ, Steart PV, Jaros E, Gatter KC. Accumulation of wild-type p53 in meningiomas. Neuropathol Appl Neurobiol 1995;21:136-42. |
|12.||Nagashima G, Aoyagi M, Yamamoto M, Yamamoto S, Wakimoto H, Ohno K, et al. P53 overexpression and proliferative potential in malignant meningiomas. Acta Neurochir (Wien) 1999;141:53-61. |
|13.||Ohkoudo M, Sawa H, Hara M, Saruta K, Aiso T, Ohki R, et al. Expression of p53, MDM2 protein and Ki-67 antigen in recurrent meningiomas. J Neurooncol 1998;38:41-9. |
|14.||Radhakrishnan K, Mokri B, Parisi JE, O′Fallon WM, Sunku J, Kurland LT. The trends in incidence of primary brain tumors in the population of Rochester, Minnesota. Ann Neurol 1995;37:67-73. |
|15.||Bollag RJ, Vender JR, Sharma S. Anaplastic meningioma: Progression from atypical and chordoid morphotype with morphologic spectral variation at recurrence. Neuropathology 2010;30:279-87. |
|16.||Wernicke AG, Dicker AP, Whiton M, Ivanidze J, Hyslop T, Hammond EH, et al. Assessment of Epidermal Growth Factor Receptor (EGFR) expression in human meningioma. Radiat Oncol 2010;5:46. |
|17.||Lusis EA, Chicoine MR, Perry A. High throughput screening of meningioma biomarkers using a tissue microarray. J Neurooncol 2005;73:219-23.17. |
|18.||Westphal M, Herrmann HD. Epidermal growth factor - Receptors on cultured human meningioma cells. Acta Neurochir (Wien) 1986;83:62-6. |
|19.||Horsfall DJ, Goldsmith KG, Ricciardelli C, Skinner JM, Tilley WD, Marshall VR. Steroid hormone and epidermal growth factor receptors in meningiomas. Aust N Z J Surg 1989;59:881-8. |
|20.||Reubi JC, Horisberger U, Lang W, Koper JW, Braakman R, Lamberts SW. Coincidence of EGF receptors and somatostatin receptors in meningiomas but inverse, differentiation-dependent relationship in glial tumors. Am J Pathol 1989;134:337-44. |
|21.||Smith JS, Lal A, Harmon-Smith M, Bollen AW, McDermott MW. Association between absence of epidermal growth factor receptor immunoreactivity and poor prognosis in patients with atypical meningioma. J Neurosurg 2007;106:1034-40. |
|22.||Kuratsu JI, Seto H, Kochi M, Ushio Y. Expression of PDGF, PDGF-receptor, EGF-receptor and sex hormone receptors on meningioma. Acta Neurochir (Wien) 1994;131:289-93. |
|23.||Manasa LP, Uppin MS, Sundaram C. Correlation of p53 and KI-67 expression with grade and subtype of ependymoma. Indian J Pathol Microbiol 2012;55:308-13. |
|24.||Devaprasath A, Chacko G. Diagnostic validity of the Ki-67 labeling index using the MIB-1 monoclonal antibody in the grading of meningiomas. Neuropathology 2003;51:336-40. |
|25.||Babu S, Uppin SG, Uppin MS, Panigrahi MK, Saradhi V, Bhattacharjee S, et al. Meningiomas: Correlation of Ki67 with histological grade. Neurol India 2011;59:204-7. |
|26.||Ghule P, Kadam PA, Jambhekar N, Bamne M, Pai S, Nair C, et al. p53 gene gets altered by various mechanisms: Studies in childhood sarcomas and retinoblastoma. Med Sci Monit 2006;12:BR385-396. |
|27.||Perry A, Stafford SL, Scheithauer BW, Suman VJ, Lohse CM. The prognostic significance of MIB-1, p53, and DNA flow cytometry in completely resected primary meningiomas. Cancer 1998;82:2262-9. |
|28.||Kamei Y, Watanabe M, Nakayama T, Kanamaru K, Waga S, Shiraishi T. Prognostic significance of p53 and p21WAF1/CIP1 immunoreactivity and tumor micronecrosis for recurrence of meningiomas. J Neurooncol 2000;46:205-13. |
|29.||Cho H, Ha SY, Park SH, Park K, Chae YS. Role of p53 gene mutation in tumor aggressiveness of intracranial meningiomas. J Korean Med Sci 1999;14:199-205. |
|30.||Lanzafame S, Torrisi A, Barbagallo G, Emmanuele C, Alberio N, Albanese V. Correlation between histological grade, MIB-1, p53, and recurrence in 69 completely resected primary intracranial meningiomas with a 6 year mean follow-up. Pathol Res Pract 2000;196:483-8. |
|31.||Aguiar PH, Agner C, Simm R, Freitas AB, Tsanaclis AM, Plese P. p53 Protein expression in meningiomas - A clinicopathologic study of 55 patients. Neurosurg Rev 2002;25:252-7. |
[Figure 1], [Figure 2], [Figure 3]
[Table 1], [Table 2], [Table 3]
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