Human Papilloma Virus Prevalence among Women with Cervical Intraepithelial
Neoplasia III and Invasive Cervical Cancer from 2001 till 2006 in BandarAbas,Iran
Sobhani (1) ,Z.Etaati (2), A.Sadeghi (1),
A.SH Jahanlou(3), M.Shiroodi (4)
To estimate the risk of Human
Papilloma Virus (HPV) infection for cervical malignancies we conducted a
case-control study in Iran.52 paraffin embedded blocks with exact
diagnosis of cervical carcinoma from years 2001 till 2006 and 52 praffin
embedded blocks of cervical tissue specimens with normal histopathology
as the control group were tested for the presence of HPV DNA using PCR
based assay.HPV DNA was found in 16 out of 52 patients (30.7%) while it
was not detected in any of the control group.
cancer is one of the most
prevalent forms of carcinoma among women world wide and accounts for
about 12% of all cancer cases among women with an incidence of more than
400/000 cases per year.Yet some studies have shown that cervical
cancer is the fifth most frequently seen cancer and the second most
common cancer in women world wide.
association between certain Human Papilloma Viruses (HPVs) and cervical
cancer is well documented and research over the past 2 decades has
convincingly revealed that HPVs are etiologically related to the
development of most cases of cervical cancer.[3-5] There is no exact
statistical data of cervical cancer prevalence among Iranian women
however cervical cancer like other forms of genital cancer is more
prevalent in the northern part of Iran including Mazandaran
province.,but generally the association between HPVs and cervical
cancer has not been studied among Iranian women except for Mazandaran
goal of our case-control study was PCR based assay of the association of
HPV DNA with cervical cancer in patients admitted to BandarAbas Shariati
hospital in Hormozgan province (south of
Iran) since years 2001 till 2006.
Specimens: We extracted
paraffin embedded blocks of cervical tissue specimens with
histopathologic diagnosis of cervical carcinoma in
the years 2001 till 2006 from archive of the pathology department
of BandarAbas Shariati hospital.After revision of corresponding slides
and assurance of consistency ,2 drops out occurred and finally 52
paraffin embedded blocks with exact diagnosis of cervical carcinoma (50
carcinomas and 2 carcinoma in situs)were entered in our study.In
parallel we selected 52 blocks of paraffin embedded cervical tissue
specimens with normal histopathology as the control group.We tried to
age match the control group with the study group.
pathology,BandarAbas University of Medical sciences 2-Department of
gynecology and obstetrics,BandarAbas University of Medical
Sciences3-Department of Educational medicine,BandarAbas University of
Medical Sciences 4-Department of genetics,BandarAbas University of
extraction: Sections of
5-10 micrometer wide were prepared from each specimen,avoiding any cross
contamination between samples (using separate disposable items such as
gloves,blades and tubes).Sections were subsequently deparaffinized by
xylene and digested using digestion buffer containing proteinase
K.,followed by extensive extraction with phenol/chloroform.The
extracted DNA was stored at 4 C until tested.
DNA quality was evaluated by PCR using primers G73/G74 that amplify a
268 bp product from the human b-globin gene.Then b-globin positive
samples were subjected to HPV PCR by GP5+/GP6+ primers for L1 open
reading frame (ORF) that amplifies a 150 bp product from the HPV L1
performed according to the procedure described by Yi Ting et al.
Samples were subsequently subjected to agarose gel electrophoresis (2%
agarose),and stained with ethidium bromide.
was found in 16 out of 52 cases (30.7%) while HPV DNA was not found in
any of the control group.(table-1)Out of 52 cases of cervical carcinoma
38 cases (73%) were squamous cell carcinomas,4 cases (7.6%) were
adenocarcinomas, 6 cases (11.5%) were poorly differentiated carcinomas
and 4 cases (7.6%) were carcinoma in situs (CIN III).At the same time 10
out of 38 (26.3%) squamous cell carcinomas, 2 out of 4 (50%)
adenocarcinomas,2 out of 6 (33.3%) poorly differentiated carcinomas and
2 out of 4 (50%) carcinoma in situs were positive for HPV DNA.(table-2)
TABLE-1: PCR RESULTS IN STUDY AND CONTROL
POORLY DIFFERENTIATED CARCINOMA
CARCINOMA IN SITU
Table-2:PCR RESULTS IN VARIOUS CARCINOMA TYPES
epidemiological studies indicate that human cancer is a multifactorial
disease,which can develop through different molecular biologic
pathways.Currently there is compelling evidence that the development of
human cervical cancer without the involvment of specific human papilloma
viruses is exceptional or impossible.The international biological
study on cervical cancer demonstrated that 92.9% of cervical cancers
from 22 countries contained HPV DNA ranging from 75 to 100%. In our
study we found that the
frequency of HPV DNA in our cervical carcinoma patients was 30.7%.This
figure is much lower than what was found in Mazandaran province(78.6%)
in Iran.Also it is lower than what was reported from
Brazil(76%),,Mozambique(92%),,and several other countries in
studies have reported difficulties in reproducing PCR results with
formalin fixed and paraffin embedded tumor tissues,therefore it is
possible that if fresh frozen tumor tissue had been examined,HPV
positive cases might have been higher because amplification could be
hampered by formalin. On the other hand our finding is much closer
to the study performed in an Islamic country like Malaysia inwhich 45.1%
of specimens were positive for HPV.Hence we conclude that before
generalizing the concept of low frequency of HPV infection in women of
Islamic countries larger number of samples from different regions need
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