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Original Article
Assessment of expression of epidermal
growth factor receptor and p53 in
meningiomas
Swetha Narla, Megha S. Uppin, M. Vijaya Saradhi1, B. P. Sahu1, A. K. Purohit1, C. Sundaram

Departments of Pathology, and 1Neurosurgery, Nizams Institute of Medical Sciences, Hyderabad, Andhra Pradesh, India

Abstract
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
Address for correspondence:
to be higher in grade I (82.93%) compared with grade II (35.71%) and grade III tumors
Dr. C. Sundaram, (20%) with an overall positivity of 60.81%. Mean p53 indices were higher in grade III (50%)
Department of Pathology, compared with grade II (39.29%) and grade I tumors (38.46%) with an overall positivity
Nizams institute of Medical Sciences, of 39.44%. Ki67 labeling index (LI) was significantly high in grade III (16.4%) compared
Hyderabad - 500 082, with grade II (6.46%) and grade I tumors (3.13%). Conclusion: EGFR expression and
Andhra Pradesh, India. Ki67 LI correlated with grade of meningioma P < 0.0001 and P < 0.0001 respectively
E-mail: challa_sundaram@yahoo.com
which were statistically significant whereas p53 expression did not correlate (P - 0.90).
Received : 13-05-2013
Review completed : 23-12-2013 Key words: Epidermal growth factor receptor, grade, Ki67 labeling index,
Accepted : 26-01-2014 meningioma, p53, tissue microarrays

Introduction morbidity and mortality. [1] Primary therapy for

Meningiomas represent about 30% of primary adult
central nervous system tumors. They are generally slow
growing tumors, but can recur and cause significant
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PMID:
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DOI:
10.4103/0028-3886.128276
meningiomas is surgery. Overall recurrence rate has
been reported to be around 20%.[2,3] 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.[4,5] Weisman, et al.[6]
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[7-9] on the expression

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Narla, et al.: Assessment of expression of EGFR and p53 in meningiomas

of EGFR assessment in meningiomas, until date there were counted and an average was taken. Comparisons of
are no studies from India. Another such marker is p53 immunoreactivity frequencies of EGFR and p53 between
which is a tumor suppressor gene located on the short tumor grades were carried out using the Chi-square test
arm of chromosome 17. The function of p53 is critical with Yates correction.
to the efficiency of many cancer treatment procedures.
Consequently, tumors which carry mutations in p53 are Results
often difficult to treat and their prognosis is poor. Studies
of p53 expression in meningiomas are limited.[10-13] This A total of 100 cases of meningiomas were diagnosed in
study was designed to determine whether meningiomas our department during the study period. Only 79 cases
express EGFR and p53 and if so, to establish a correlation were included in the present study where sufficient
between the histopathologic grade of these tumors and material was available in the blocks. These included 30
the degree of expression. 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).
Materials and Methods There were 46 grade I meningiomas [Figure 1a] (58.23%), 28
grade II meningiomas [Figure 2a] (35.44%) and 5 grade III
This was a retrospective study. All cases of meningiomas meningiomas [Figure 3a] (6.33%). There were 70 (88.61%)
diagnosed in the year 2010 in the department of intracranial and 9 (11.39%) spinal meningiomas. Among
pathology were included in the study. All hematoxylin 79 cases of meningiomas, transitional meningiomas were
and eosin stained slides were reviewed and graded the most common subtype both in intracranial (31/70;
according to World Health Organization (WHO) criteria 44.29%) and spinal (9/9; 100%) location. Table 1 shows
2007. [1] Presence or absence of brain invasion was the demographic data, clinical features and IHC results
noted. The best section which was free from necrosis or of meningiomas.
hemorrhage was selected for tissue microarray (TMA).
Representative areas from each section were marked on Immunohistochemical analysis
the tissue block from which 3 cores were extracted. Two The final analysis for IHC was carried out on EMA
TMA blocks were prepared from the paraffin blocks by (79 cases), Vimentin (79 cases), EGFR (74 cases), p53
manual method. One TMA block consisted of 120 cores (71 cases) and Ki67 LI (79 cases) as some of the cores
from 40 blocks and the other TMA block consisted of showed edge artefacts and some were lost in processing.
117 cores from 39 blocks.
IHC with EMA and Vimentin
Immunohistochemistry (IHC) EMA and Vimentin immunopositivity was seen in all
Immunohistochemical analysis with epithelial membrane the cases and in all grades of meningiomas (79/79; 100%
antigen (EMA), Vimentin, Ki67 (ready to use Biogenex, positivity).
USA), EGFR (monoclonal mouse antibody, 1:100,
Biogenex, USA) and p53 (monoclonal antibody, 1:50, IHC with EGFR
DAKO, USA) was performed by horseradish peroxidase EGFR expression was higher in grade I tumors (82.93%)
polymer technique on the TMA slides. Cores were [Figure 1b] when compared with grade II (35.71%)
recorded as non-informative if the tissue was lost in
processing/there was no recognizable tumor in the Table 1: Summary of demographic data, clinical features and
core/or there were extensive staining artefacts (e.g. immunohistochemical results of meningiomas
inappropriate staining of collagen or tissue edges WHO grade Grade I Grade II Grade III
and tissue creases in a specimen with minimal tissue (%) (%) (%)
No. of cases 46/79 (58.23) 28/79 (35.44) 5/79 (6.33)
retained). EGFR expression was considered positive if it
Male: Female 16:30 12:16 2:3
stained either cytoplasm/cell membrane and was scored M: F 1:1.88 1:1.33 1:1.5
according to the percentage of stained cells. Mean age in years at 46.52 45.25 49.60
operation
Scoring for p53 nuclear staining was based on a Most common location Spinal Frontal Frontal
of tumor
four-point scale: no staining 0; <5% of nuclei with No. of tumors (%) 9 (19.57) 6 (21.43) 3 (60)
positive staining 1; 5-30% of nuclei with positive staining No. of intracranial 37/46 (80.43) 28/28 (100) 5/5 (100)
2; and 30% of nuclei with positive staining 3. For the tumors
purpose of analysis, all p53 scores of 0 and 1 were No. of spinal tumors 9/46 (19.57) - -
EGFR expression 34/41 (82.93) 10/28 (35.71) 1/5 (20)
considered as negative, whereas the scores of 2 and 3 p53 expression 15/39 (38.46) 11/28 (39.29) 2/4 (50)
were considered as positive. For counting Ki67 labeling Ki67 LI meanSD 3.13%3.04 6.46%5.37 16.4%8.62
index (LI) and p53, 100 nuclei/core were counted and an SD - Standard deviation, WHO - World Health Organization,
average was taken and for counting EGFR, 100 cells/core EGFR - Epidermal growth factor receptor, LI - Labeling index

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Narla, et al.: Assessment of expression of EGFR and p53 in meningiomas

[Figure 2b] and grade III tumors (20%) [Figure 3b] 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%)
a b
[Figure 1c] compared with grade II (39.29%) [Figure 2c]
and grade III meningiomas (50%) [Figure 3c] 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 1d] to grade II (6.46%) [Figure 2d] to
grade III tumors (16.4%) [Figure 3d]. These values were
statistically significant (P 0.0001).
c d
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)

Discussion

WHO has recognized three grades in meningiomas

which helps to predict the clinical behavior of these
tumors. [1] 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
a b
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.[14]
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.,[15] grade II and grade III meningiomas were
more common in men.
c d
Despite many recent modifications in the histological Figure 2: Grade II meningioma (a) grade II meningioma-high cellularity
with loss of architectural pattern, mitosis (H and E, 100). (b) Tumor cells
classification of meningiomas, there is still considerable stained positive with epidermal growth factor receptor (100). (c) Nuclear
variability in the biological behavior within each of the staining with p53 (100). (d) Nuclear staining with Ki67 (100)
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 a b
meningiomas as defined by the WHO 2007 criteria.[7,16-18]
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.,[16] EGFR expression was noted in c d
86% of all meningiomas. Lusis et al.[17] showed that EGFR Figure 3: Grade III meningioma (a) grade III (rhabdoid) meningioma (H
and E, 100). (b) Tumor cells stained positive with epidermal growth
expression was seen in 88% of the incidentally detected factor receptor (100). (c) Nuclear staining with p53 (100). (d) Nuclear
meningiomas and only 20% of the grade I, 61% of grade staining with Ki67 (100)

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Narla, et al.: Assessment of expression of EGFR and p53 in meningiomas

II and 44% of grade III meningiomas demonstrated Table 2: Summary of immunohistochemistry results with EGFR
immunoreactivity for EGFR (P - 0.032). In other studies according to different studies
EGFR has been found in nearly 100% of meningioma Author[ref] Grade I (%) Grade II (%) Grade III (%)
specimens in tissue culture.[18-20] Wernicke et al. (n=85)[16] 52/57; 92 20/23; 87 1/5; 20
Lusis et al. (n=33)[17] 1/5; 20 11/18; 61 5/10; 50
Reports vary on EGFR expression in meningiomas, Andersson et al. (n=26)[7] 15/26; 58 - -
Smith et al. (n=61)[21] 16/33; 48 10/21; 48 3/7; 42
which range from 33% to 100% respectively [Table 2].
Present study (n=74) 36/41; 82.93 10/28; 35.71 1/5; 20
In majority of studies, the expression ranged between
EGFR = Epidermal growth factor receptor
50% and 60% respectively.[17-20,22] Different techniques
were used in different studies, which may be the cause
for this variation in expression rates. Andersson et al.[7] Table 3: Summary of previous studies of correlation of p53 with
utilized immunohistochemical and quantitative real-time histological grade
polymerase chain reaction to identify EGFR expression Author[ref] Correlation of p53 with histological grade
in 26 cases. They did not identify an association between Wang et al.[10] Statistically significant
Nagashima et al.[12] Statistically significant
expression level and outcome, but their study included
Ohkoudo et al.[13] Statistically significant
only cases of grade I meningiomas. Manasa et al.[23] also Aguiar et al.[31] Statistically significant
carried out TMA IHC technique similar to our study but Ellison et al.[11] No significant correlation
on the correlation of p53 and Ki67 expression with grade Present study No significant correlation
and subtype of ependymomas.
grade. The physiological significance of the role of EGFR
The present study was conducted with the objective to
in human meningioma is unknown at present, but
evaluate the expression of p53 and Ki67 in different grades
knowledge of factors which regulate tumor growth can
of meningiomas. Chacko et al.[24] showed that grade II
lead to a possible treatment when tumors are inoperable.[15]
tumors had a higher MIB-1 LI than grade I tumors. The
Application of EGFR targeted therapies to meningiomas
MIB-1 LI used in conjunction with histological features can
requires further trials to be conducted. By detecting
help in making a recommendation regarding potentially
EGFR mutations, meningiomas could become eligible for
aggressive behavior in meningiomas. Babu et al.[25] studied
targeted therapy. Immunohistochemical evidence of EGFR
Ki67 LI in various histological subtypes and grades of
expression is a prognostic indicator in meningiomas.
meningioma and correlated it with various parameters for
recurrence. They showed that high Ki67 LI correlated with
higher grade of meningioma. Pratibha et al.[26] provided Acknowledgments
evidence of p53 alteration through allelic deletion that
are common primary somatic mutation events which The authors gratefully acknowledge Dr. Sanjay Navani for
helping in preparation of tissue microarray blocks.
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How to cite this article: Narla S, Uppin MS, Saradhi MV, Sahu BP,
Purohit AK, Sundaram C. Assessment of expression of epidermal
growth factor receptor and p53 in meningiomas. Neurol India
2014;62:37-41.
Source of Support: Nil, Conflict of Interest: None declared.
41
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