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www.ijpbs.net
Internationally indexed journal
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The “International Journal of Pharma and Bio Sciences” (IJPBS) is an international journal in English
published quarterly. The aim of IJPBS is to publish. peer reviewed research and review articles rapidly
without delay in the developing field of pharmaceutical and biological sciences
Indexed in Elsevier Bibliographic Database
(Scopus and EMBASE)
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Impact factor 0.67*
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Int J Pharm Bio Sci 2014 April ; 5 ( 2 ) : (B) 253 - 260
Research Article
Pathology
International Journal of Pharma and Bio Sciences
ISSN
0975-6299
HISTOPATHOLOGICAL STUDY OF PROSTATIC LESIONS AND
ASSESSEMENT WITH AGNOR INDEX
K.SUBATHRA*1 AND N.SANGEETHA2
1Department of pathology, Madurai medical college, Madurai, Tamil Nadu, India.
2Department of pathology, Stanley medical college, Chennai, Tamil Nadu, India.
ABSTRACT
Interpretation of prostatic biopsies has been a continuous problem for practicing
pathologist. This current study aims at the analysis of histopathological features of
various non neoplastic and neoplastic lesions of the prostate including the grading of
malignant lesions and evaluation of role of AgNOR index in different benign,
premalignant and malignant lesions of prostate. One hundred and eight transurethral
resection of prostate specimens were studied with haematoxylin and eosin and AgNOR
staining. Benign prostatic hyperplasia was the most common lesion affecting the
prostate in elderly. High grade prostatic intra epithelial neoplasia had strong association
with prostatic carcinoma. Among the malignant lesions of the prostate, primary prostatic
adenocarcinoma was the commonest. According to Gleason Grading system higher
grades were more commonly observed as the predominant pattern. Mean AgNOR
counts were higher in malignant lesions when compared with the benign lesions.
KEYWORDS: Benign prostatic hyperplasia, Prostatic adenocarcinoma, AgNOR
K.SUBATHRA
Department of pathology, Madurai medical college, Madurai, Tamil Nadu, India.
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Int J Pharm Bio Sci 2014 April ; 5 ( 2 ) : (B) 253 - 260
INTRODUCTION
Prostatic diseases, benign and malignant are
collectively responsible for significant morbidity
and mortality in men throughout the world. Only
three pathologic processes affect the prostate
gland with sufficient frequency to merit
discussion
namely
inflammation,
benign
prostatic hyperplasia (BPH) and tumors. Benign
prostatic hyperplasia (BPH) is common in
elderly men and It is characterized by nodular
proliferation of both glandular and stromal
components.. Prostate cancer is now the sixth
most common cancer in the world. Prostatic
adenocarcinoma is the second most common
cause of cancer mortality in men next to lung
cancer. Prostatic cancers are diagnosed by fine
needle aspiration cytology, needle biopsies,
transurethral resection of prostate (TURP) and
prostatectomy. Various types of difficulties have
been encountered while diagnosing and typing
prostatic carcinoma and premalignant lesions
especially in TURP chips where there is loss of
orientation and coagulation of tissue during
cauterization. Prostatic lesions on routine
haematoxylin & eosin staining sometimes cause
diagnostic difficulties in identifyingpremalignant
and
malignant
conditionsAn
important
diagnostic criterion in the differentiation is the
loss of basal cell layer in adenocarcinoma and
its presence in the benign lesions. Several
immunohistochemical markers have been used
to stain the basal cells of prostate like High
molecular weight cytokeratin (HMWCK), p63
etc. Proliferative markers like silver staining
nucleolar
organizer
regions
(AgNOR),
proliferating cell nuclear antigen (PCNA) are of
great help in differentiating benign premalignant
and malignant lesions. This current study aims
at analysis of histopathological features of
various non neoplastic and neoplastic lesions of
the prostate including the grading of malignant
lesions and evaluation of role of proliferative
markers in different benign, premalignant and
malignant lesions of prostate. Histological
typing, grading and staging of prostatic
carcinoma are vital in planning the treatment
strategies and predicting the survival rate.
MATERIALS AND METHODS
This present study is a prospective study
undertaken in the department of pathology,
Madurai Medical College, Madurai, during the
period of May 2009 to July 2011. This study
was conducted on 108 prostatic specimens of
which 56 specimens were from Government
Rajaji Hospital, Madurai and 52 were from
Madurai Kidney centre, Madurai. All the 108
specimens received were Trans Urethral
Resection of Prostate (TURP) specimens
ranging in volume from 1 to 10 grams. These
were fixed in 10% neutral buffered formalin for
12 hours. After adequate fixation, the
specimens were submitted for processing until
four cassettes were filled. Tissue processing
was done with automated tissue processor and
sections were made manually with microtome of
thickness 2-4 microns. Staining was done with
routine haematoxylin and eosin and examined
under light microscope. Silver staining of
nucleolar organizer region (AgNOR) method of
Smith and Crocker was done for all the cases
taken for study excluding the cases of
Leiomyosarcoma of prostate and contiguous
spread of rectal adenocarcinoma to prostate. All
the slides were examined under 100X oil
immersion objective with 10X eye piece. One
hundred lesional nuclei of epithelial cells were
taken at random for the counting procedure.
Careful focusing allowed the nucleolar organizer
regions (NOR) to be visualized as black dots
arranged both in clusters and clumps and as
individual “satellites” within the cell nucleolus.
The NOR dots were counted per nuclei and an
average count was noted.
RESULTS
Benign prostatic hyperplasia was the most
common type of lesion – 96 cases (88.89%),
followed by malignant lesion – 10 cases
(9.26%) and inflammatory lesions – 2 cases
(1.85%). The incidences of various prostatic
lesions are illustrated in Graph1. Among the 98
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Int J Pharm Bio Sci 2014 April ; 5 ( 2 ) : (B) 253 - 260
benign lesions, youngest case reported was 41
years old and oldest was 81 years. The mean
age group for benign lesions was 63.44. Among
the 10 malignant lesions, youngest case
reported was 50 years old and oldest was 77
years. The mean age group for malignant
lesions was 63.90. Two cases (1.85%) showed
features of granulomatous prostatitis. (Figure 1).
Out of all the prostatic lesions studied, Nodular
hyperplasia constituted the bulk of the lesions
[88.89 %
( 96 cases)]. Foci of low grade
PIN were identified in12 cases (11.11%). Foci of
High grade PIN was identified in three cases
(2.78%). Among the three cases one was BPH
and other two were adenocarcinoma. (Figure 2)
In our prospective study, malignant lesions
constituted the second most common pathology
of prostate. This study included a total of 10
cases of malignant lesions of prostate.
Adenocarcinoma was the most common [7.41%
(8 cases)] type of primary carcinoma
encountered. Others were Leiomyosarcoma of
prostate invading the bladder [1 case (0.93%)]
and the Rectal adenocarcinoma invading the
prostate [1 case (0.93%)]. All the cases of
primary prostatic carcinoma were graded using
Gleason scoring system. Primary grade was
assigned to dominant pattern and secondary
grade to second dominant pattern and then the
two numeric scores were added to obtain
combined Gleason score. In tumors with one
pattern of arrangement, the number was
doubled. (Table 1) (Figure 3, 4, 5) Mean
AgNOR count was higher in malignant lesions
when compared to benign lesions. (Figure 6, 7,
8), (Table 2. One way Analysis Of Variance test
was used to assess the statistical difference
between benign, premalignant and malignant
lesions. The differences in the mean value
between benign and pre malignant lesions were
statistically significant. (P = < 0.001). The
differences in the mean value between benign
and malignant lesions were statistically
significant. (P = < 0.001).
Graph 1
Incidence of various prostatic lesions
INCIDENCE OF VARIOUS PROSTATIC LESIONS
9.26
1.85
BPH
Malignant
Inflammatory
88.89
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Int J Pharm Bio Sci 2014 April ; 5 ( 2 ) : (B) 253 - 260
Table 1
Gleason’s grading for adenocarcinoma
S.NO
PATH NO
HPE DIAGNOSIS
1
2
3
4
5
6
7
8
3667/09
3909/09
2674/10
36/09
50/09
78/10
25/11
46/11
Adenocarcinoma
Adenocarcinoma
Adenocarcinoma
Adenocarcinoma
Adenocarcinoma
Adenocarcinoma
Adenocarcinoma
Adenocarcinoma
GLEASON’S
GRADE
3+3
2+2
3+4
1+2
1+4
3+2
2+3
3+4
GLEASON’S
SCORE
6
4
7
3
5
5
5
7
AgNOR
count
5.1
4.5
5.5
4
4.5
4.8
4.9
5.2
Table - 2
The mean AgNOR count in various prostatic lesions
Lesions
Granulomatous prostatitis
BPH with or without prostatitis
BPH with LGPIN
BPH with HGPIN
Adenocarcinoma
No of cases
2
83
11
1
8
Mean AgNOR count
1.9
1.43
2.08
3.9
4.81
Table 3
Comparison of mean AgNOR count in various studies
Studies
13
Sakr, W. A
11
Asim Kumar Manna
Present study
BPH
1.836
1.3
1.44
PIN
3.129
4.7
2.23
Prostatic adenocarcinoma
4.737
4.91
4.81
Figure 1
Nonspecific granulomatous prostatitis showing foamy
histiocytes and multinucleated giant cells (H & E 400X) (21/10).
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Int J Pharm Bio Sci 2014 April ; 5 ( 2 ) : (B) 253 - 260
Figure 2
Foci of High grade PIN showing epithelial cell crowding and stratification
in tufting pattern with enlarged nuclei and prominent nucleoli. (H&E 400X)
Figure 3
Figure 4
Figure 5
Figure 3: Prostatic adenocarcinoma Gleason’s pattern 2 –loosely packed single glands with
irregular edges. (H&E 100X) (78/10), Figure 4: Prostatic adenocarcinoma Gleason’s pattern 3 –
scattered single glands. (H&E 100X) (78/10), Figure 5: Prostatic adenocarcinoma Gleason’s
pattern 4 – fused infiltrating glandular pattern. (H&E 100X) (2674/10)
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Int J Pharm Bio Sci 2014 April ; 5 ( 2 ) : (B) 253 - 260
Figure 6
Figure 7
Figure 8
Figure 6: BPH showing occasional AgNOR dots per nuclei. (AgNOR stain 1000X) (2116/10),
Figure 7: High grade PIN showing two to three AgNOR dots per nuclei. (AgNOR stain 1000X)
(103/10), Figure 8: Prostatic adenocarcinoma showing numerous AgNOR dots per nuclei.
(AgNOR stain 1000X) (2674/10)
DISCUSSION
The incidence of BPH increases with age. BPH
is seen in 20% of the men at 40 years of age, a
figure that increases to 70% by age of 60 and to
90% by age of 80. In the present study 108
cases of TURP specimen were examined.
Benign prostatic hyperplasia was seen in 96
cases. Highest incidence of nodular hyperplasia
was noted in the 7th decade1. In a classic paper
by Berry et al2, the prevalence of BPH ranged
from 8% for men in their 30s to 88% for men
over 80s. In another classic paper Issac and
Coffey3 compared the prevalence of BPH by
age in autopsy studies from various countries.
This study demonstrated relatively similar
prevalence of BPH across a spectrum of
countries and ethnicities. In our study also the
incidence of BPH increases with age reaching
maximum in 7th decade. The decline in the
number of cases beyond the age of 80 years
may reflect the average life span of people in
our country. Prostatic Intra Epithelial Neoplasia
(PIN) has high predictive value as a marker for
adenocarcinoma. This is particularly true for
high grade PIN; if this lesion is identified; close
surveillance and follow up biopsy are indicated.
The frequency of HGPIN in transurethral
resection of the prostate specimens is between
2.3% and 4.2 %4. The present study was
comparable well with the above study, stating
2.78% prevalence of HGPIN in all TURP
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Int J Pharm Bio Sci 2014 April ; 5 ( 2 ) : (B) 253 - 260
specimens. The frequency of HGPIN in
prostates involved with cancer is significantly
increased when compared with the cancer free
prostates. The incidence of high grade PIN is
low in our study because all the specimens
were TURP which does not have enough
material compared to whole prostate specimen
examined in other studies. In a study by Pacelli
A and Bostwick DG5 the incidence of high grade
PIN in TURP specimens without carcinoma was
2.8%. The incidence of PIN in prostatic
malignancy, as quoted in the literature, varies
from 33% to 100%, depending on the nature of
specimen.6,7 . In our study, we observed high
grade PIN in 25% of the carcinomas.
In this study, malignant lesions account
for 9.26% (10 cases) of cases. Among the
malignant lesions incidence of primary prostatic
adenocarcinoma is high [8cases (80%)].
Prostate cancer is now the sixth most common
cancer in the world. The prevalence of prostatic
adenocarcinoma in this study is 7.41% (8
cases). All these cases were incidental
adenocarcinoma which were identified in
transurethral resection of the prostate (TURP)
done for BPH. According to WHO study, when
TURP is done without clinical suspicion of
cancer, prostate cancer is incidentally detected
in approximately 8- 10% of the specimens8,
which is in correlation with our study. The risk of
prostate cancer rises very steeply with age.
Worldwide, about three-quarters of all cases
occur in men aged 65 or more8. In our study
also maximum numbers of cases were found in
the age group of 60-69 years1. Gleason scoring
system is the most widely used and officially
recommended system for scoring prostatic
adenocarcinoma9,10 . Gleason score correlates
with prognosis after radical prostatectomy and
with outcome following radiotherapy. Gleason
grade on biopsy can influence mode of
treatment. We applied Gleason grading system
for all adenocarcinoma (Table2) Tumor
differentiation and proliferative activity are
important predictors of biological behavior.
While routine histological evaluation is fairly
adequate to assess differentiation, tumor
proliferative activity is difficult to measure. Silver
staining for nucleolar organizer regions
(AgNORs) is reported to be helpful for
assessing tumor proliferation. AgNOR value is
significantly higher in Prostatic carcinoma than
BPH &PIN. It is also significantly higher in PIN
than BPH. The mean AgNOR value is higher in
adenocarcinoma with high gleason score than
the lower gleason score. (Table2) Our study
showed identical results with the study done by
Asim Kumar Manna et al11. Kawase12 found
AgNOR counts were higher in carcinoma (4.2+/1.57) than in benign lesions (1.9 +/- 0.24). Wael
A sakr13 found that AgNOR study is helpful in
assessing tumor proliferation (Table 3).
CONCLUSION
We conclude that mean AgNOR counts are
significantly higher in adenocarcinoma when
compared to benign and premalignant lesions
and correlated with histological grade of the
tumor. The proliferative markers have a
significant role in the diagnosis of prostatic
lesions especially which fall in the premalignant
category and create difficulty in the diagnosis
by routine histopathological study.
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