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Iraqi Journal of Genetic

2008

Chromosome 5 deletion in two families patients with familial


adenomatous polyposis
Nahi Yousif Yaseen*
* Iraqi Centre for Cancer and Medical Genetics Research (ICCMGR), Baghdad, Iraq.

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.

Polyps originating in the epithelium of


colorectum constitute frequent findings in several
countries. The distribution of adenomas in the
colorectum is varied depending on the type and
size of the adenomas. The role of heritable factor
in the genesis of colorectal cancer (CRC) is not
well understood. However, CRC is found is to be
occur in some inherited syndromes such as familial
polyposis coli (FPC) or familial adenomatous
polyposis (FAP) and Gardner syndrome (1, 2, 3). It
was reported that FAP is inherited as an autosomal
dominant trait (4) and affects about one in ten
thousands individuals. Several evidences have
been reported by a number of observations which
established that most if not all CRC arise from
adenomas (5, 6, 7, 8, 9, 10, 11).
Cytogenetic studies have shown variable
chromosomal changes in the adenoma cells (12,
13, 14, 15, 16, 17). In another hand chromosomal
findings in peripheral blood of patients with FAP
showed interstitial deletion in the long arm of
chromosome 5 (18, 19, 20, 21, 22, 23).
Linkage study in FAP families mapped the
FPC gene to the area 5q15 to 5q22 (24). Further
studies revealed a possible association of an
interstitial deletion in chromosome 2 (2q14.3-21.3)
in the blood of FAP and Gardner syndrome
patients (25, 26).

Patients and Methods:


Subjects:
Two very close related families (Fig.1) were
studied and their members agreed to undergo the
tests required for this study. Family 1 consists of a
mother, dead (car accident) father, three daughters
and one son. The two oldest daughter were
mentally retarded, and died when they were six
years and eight years old. The youngest daughter
(18 years old) was mentally retarded, whereas their
only brother seemed to be normal. Family 2
consists of a mother, dead father (due to an
operable colon cancer with polyposis), one
daughter, and four sons. Two of these sons were
mentally retarded identical twins (12 years old).
Blood sample:
Venous blood sample (5 ml) was drawn from
all alive members of both families and put in a
sterile tubes containing heparin.
Sigmoidoscopy:
All members of both families (12 persons)
were subjected to sigmoidoscopy for polyposis
assessment.

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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The cell suspension was then centrifuged at


1100 rpm for 5 minutes, and re-suspended in
appropriate amount of a freshly made fixative to
make the suspension thin cloudy. Cell suspension
(2-3 drops) was dropped onto wet, chilled, grease free slides and allowed to air dry at room
temperature. Three days old slides were incubated
at 80 C for three hours. Trypsin solution was
spread over slide for 20 seconds and then washed
by Sorenson's buffer and stained by Giemsa stain
for two minutes. Slides were left to air dry and
then examined. The description of chromosomal
rearrangements was performed according to the
International System for Human Cytogenetics
Nomenclature (27).

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.

Iraqi Journal of Genetic

monozygotic twins (No.7, 8) in family 2 and in the


mentally retarded girl (No.4) in family 1. They
showed interstitial deletion of chromosome 5
(5q23) in all karyotyped cells (Fig.2, 3, and 4). All
other members of both families revealed normal
karyotype.

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

This study has shown chromosome 5


aberrations in the peripheral blood lymphocytes of
several members of very close related families
who exhibited colonic polyps. Another interesting
thing in this study that the two individuals of the
monozygotic twins showed the same structural
abnormality of chromosome 5 and their father died
in his fifties of colon cancer with polyposis, such
study has not been reported before.
Despite high numbers of well spread nicely
banded metaphases were fully analysed, all
members of both families showed normal
karyotypes except the monozygotic twins (No.7, 8)
and their cousin girl (No.4). They revealed
consistent abnormality of chromosome 5. This
abnormality represented by interstitial deletion of
the long arm of chromosome 5 (5q23). However,
interstitial deletion of 5q21 followed by inversion
of 5q23 cannot be rule out, but the first suggestion
is most acceptable. These results encouraged us to
subject all members of both families to
sigmoidoscopy and cytogenetic analysis for their
blood lymphocytes. All members except No4, 7,
and 8 were polyps free and showed normal
chromosomal complements. Interstitial deletion of
the long arm of chromosome 5 was reported in
blood lymphocytes of patients with FAP and
Gardner syndrome (18, 19, 20, 21, 22). Molecular
studies have mapped the gene for FAP to 5q215q22 and this FAP locus may encode for a tumour
suppressor gene (24,28). Deletion 5q was also
detected in several polyps (29) and in colorectal

Iraqi Journal of Genetic

suspected cases in order to get early detection for


patients who are prone to develop cancer (40).
Further studies are needed to follow up such
patients and to screen further families for such
inherited diseases. These findings showed that
lymphocyte chromosomal analysis may aid in
identifying individuals who are genetically
susceptible and at high risk of developing CRC.

Dysplasia in a Patient with Attenuated Familial


Adenomatous Polyposis Syndrome. J.C.O 26:
3641-3642
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