Professional Documents
Culture Documents
2008
Abstract:
Karyotypic analysis by using G-banding procedure was performed on blood lymphocytes from all
alive members of two very close related families. All members were also subjected to sigmoidoscopical
examination. Three members from both families (identical twins and their cousin girl) showed colonic
polyposis and revealed the same interstitial deletion of long arm of chromosome 5q23 in their
peripheral blood lymphocytes. This study suggests that abnormalities of certain gene(s) on 5q may
have a specific role in tumourgenesis of colon cancer in human.
Introduction:
It seems that colonic polyps in Iraq has not
been cytogenetically studied yet. Therefore, In this
study chromosomal analysis has been performed
on the members of two related families some of
them showed FAP.
Correspondence to:
Dr. Nahi Y. Yaseen, ICCMGR, Al-QadisiyaQ., Sec.603, Street
23, Behind Al-Yarmook Hospital, Baghdad, Iraq.
Mob. No.: +9647901475964
E-mail: nahiyaseen@yahoo.com
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Cytogenetics:
A half ml of heparinised whole blood was
cultured in a sterile universal containing 10 ml of
TC199 medium enriched with 20% pooled AB
blood group human plasma and 2% PHA, and
incubated at 37C. Methotrexate (0.1 ml) was added
to the culture to give a final concentration of 10 M
after 48 hrs incubation, and the culture was
incubated for a further 17 hrs. The medium
containing methotrexate was then replaced by 10
ml fresh medium supplemented with thymidine
and incubated for a further 6 hrs. Colcemid (0.2
ml) was added to the culture 15 minutes before the
end of culture time and incubated for 15 minutes.
The culture was transferred to a centrifuge tube
and spun at 1100 rpm for 10 minutes. Blood cells
were suspended in a pre-warmed (37C) hypotonic
solution (0.0375M KCl), put in a water bath (37C)
for 20 minutes and centrifuged at 1100 rpm.
The cells then were fixed three times in a
freshly made 3:1 methanol - glacial acetic acid
(v/v). After final fixation process, 5 ml of a freshly
made fixative was added to the cells, mixed well
and stored at -20 C for at least two hours.
Results:
Sigmoidoscopy:
The monozygotic twins (No.7,8) from family 1
and their cousin mentally retarded youngest
girl (No.4) were found to have many variable
sizes colonic polyps. However, the other
members of both families were polyps free (Fig.1).
Fig.1: Pedigree of two very close related families illustrating the members with FAP.
Cytogenetics:
For each individual in both families at least 50
well spread nicely banded metaphases were fully
karyotyped. The modal number of chromosomes in
all cases was normal (46). A chromosome
structural abnormality was detected in the
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Discussion:
cancer cases (30, 31). Moreover, allelic deletion of
chromosome 5 was observed in sporadic and
inherited CRC (8, 15, 32, 33, 34, 35). It was
hypothesized that FAP locus in colorectal
neoplasia is directly involved in inducing the wide
spreading epithelial hypoproliferation that precede
the formation of adenoma in patients with polyps
(36). Thus FAP locus may normally act as a
negative
regulator
of
colonic
epithelial
proliferation. Mutation of FAP gene may cause
benign adenomatosis but not carcinoma (37),
whilst changing of chromosome 5 gene is not
enough for cancer initiation; it may cooperate with
genetic abnormalities at other loci for tumour
progression. However, allelic deletion of 5q
appears to be capable of mediating steps in the
progression from normal to malignant colonic
mucosa.
It was suggested that sporadic and inherited
cancers could both result from a mutation in the
same allele (38). In another hand Vogelstein and
his colleagues (39) showed that allelic loss of 5q
increased with increase progression of adenoma to
carcinoma.
In this study, three members from two related
families showed polyposis and the identical twins
father died of CRC with polyposis, this may give
an impression that they suffer from FAP.
Moreover the grandma of the twins revealed that
their granddad died also from abdomen pain but
had not been specifically diagnosed at that time.
Genetic counseling may be very helpful for
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