Content uploaded by S.R. Das
Author content
All content in this area was uploaded by S.R. Das on Oct 15, 2016
Content may be subject to copyright.
Naik et al., IJPSR, 2016; Vol. 7(1): 206-220. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 206
IJPSR (2016), Vol. 7, Issue 1 (Research Article)
Received on 08 July, 2015; received in revised form, 04 September, 2015; accepted, 06 November, 2015; published 01 January, 2016
PHILADELPHIA CHROMOSOME IN PRETHERAPEUTIC CASES OF CHRONIC
MYELOGENOUS LEUKEMIA
Manoj Kumar Naik 1, Dharma Niranjan Mishra 1, Subhrajita Rout *2, Priyambada Panda 2,
Sitansu Kumar Panda 2, Saurjya Ranjan Das 2, Tapaswini Mishra 2and Chinmayi Mohapatra 3
Department of Anatomy 1, VSS Medical College, Burla, Odisha, India.
Department of Microbiology 2, IMS and SUM Hospital, Siksha ‘O’ Anusandhan University, K8, Kalinga
Nagar, Bhubaneswar-751003 India.
Department of anatomy 3, SCB medical, Cuttack, Odisha, India.
ABSTRACT: Chronic myeloid leukemia (CML) is a clonal myel0 proliferative
disorder due to neoplastic transformation of myeloid stem cells. The characteristic
Philadelphia Chromosome translocation t(9:22) (q 34: q11) juxta poses the c-abl
oncogene from chromosome 9 with break point cluster region (bcr) on chromosome
22 resulting in the generation of aberrant bcr/abl transcripts. The abnormal bcr/abl
tyrosine kinase gene product has enhanced activity compared to the wild type c-abl
tyrosine kinase and is believed central to the pathogenesis of CML. Most of the
patients in chronic phase who are treated with cytotoxic agents eventually develop a
fatal blast phase that is the end stage in which the patient usually fails to respond
combination chemotherapy and only treatment remain after that is allogenic bone
marrow transplantation. Since the philadelphia chromosome is the most consistant
abnormality in CML patients and as it possess both diagnostic as well as prognostic
importance, it’s detection is mandatory to establish the diagnosis of CML and for
planning of chemotherapy. Our study is a preliminary work to re-establishing the
finding of previous workers and to detect out any other associated or new reccuring
chromosomal anomalies in CML patients. The bone marrow samples of 89 CML
patients (54 males & 35 females) diagnosed clinically and pathologically were
collected in heparinised RPMI-1640 culture media aseptically with informed consent
from patients and their guardian in Hematology section of pathology Dept. during
the time period from Nov-2006 to May 2008. But the most common additional
structural chromosomal aberration found here was t(15:17), which is relatively rare
in available literature. To establish this as a newly recurring additional chromosomal
aberration in CML patients, large database analysis is required.
INTRODUCTION: Chronic Myelogenous
Leukemia (CML) is a clonal expansion of
haematopoeitic progenitor cells characterised
clinically by myeloid hyperplasia, leukocytosis
with basophilia and splenomegaly.
QUICK RESPONSE CODE
DOI:
10.13040/IJPSR.0975-8232.7(1).206-20
Article can be accessed online on:
www.ijpsr.com
DOI link: http://dx.doi.org/10.13040/IJPSR.0975-8232.7 (1).206-20
The cytogenetic hall mark of CML is the
philadelphia (Ph) translocation t(9:22) (q34:q11),
resulting in P 210 BCR-ABL which is essential for
the development of the disease. In 1960, Nowell
and Hungerford described the characteristic
philadelphia chromosome associated with CML 1.
The first recurrent chromosomal abnormality
associated with a specific human malignancy. They
reported that myeloid cells from patients with CML
showed a deletion of long arm of chromosome 22.
Subsequently, the translocation t(9:22) was
defined. Present work is a cytogenetic study on
CML patients to detect philadelphia chromosome
Keywords:
Philadelphia chromosome,
CML, leukemia, karyotyping
Correspondence to Author:
Ms. Subhrajita Rout
Department of Microbiology,
IMS and SUM Hospital, Siksha ‘O’
Anusandhan University, K8, Kalinga
Nagar, Bhubaneswar-751003, India.
E-mail: subhramicrobiology@gmail.com
Naik et al., IJPSR, 2016; Vol. 7(1): 206-220. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 207
and other associated structural and numerical
chromosomal aberrations, before any therapy
introduced to them. For this bone marrow samples
were collected and analysed by direct harvesting
and giemsa banding. Karyotyping was done and the
findings were compared and correlated with that of
previous workers.
MATERIAL & METHODS:
The study was carried out at S.C.B Medical
College, Cuttack in the Dept. of Anatomy in
collaboration with Hematology section of
Pathology, Dept. for sample collection and
Telemedicine Dept. for microphotography. In the
present study, bone marrow from CML patients
were sampled for analysis. The samples were
collected from both male and female patients
coming to Hematology Section of Pathology
Department of S.C.B Medical College, Cuttack.
Patients within the age group of 2 - 70 yrs have
been analysed for this study.
Criteria for selection of Patients:
The patients were selected on the basis of clinical
finding such as hepatosplenomegaly, unexplained
anemia, and pyrexia of unknown origin along with
pathological feature of leukocytosis, basophilia and
presence of blast cells in peripheral Smear.
Detailed inquiry about family history was done and
informed consent was taken from the patients for
cytogenetic analysis.
Cytogenetic Study:
The following reagents were used for the present
study.
Anticoagulant: Heparin solution was used as
anticoagulant in the dose of 100 IU per ml of bone
marrow. Heparin was added to the culture medium
and mixed thoroughly before sample collection.
Culture Media: Heparinized RPMI 1640 culture
media with L. glutamine was used for sample
collection. The necessary micronutrients and
essential amino acids required for the cell survival
are present in this culture medium.
Spindle Inhibitor: Colchicine was used as spindle
inhibitor which arrest the mitosis at metaphase
stage. This result in accumulation of chromosomes
in rappidly dividing cells. For better results 0.005
mg of colchicine was added in 10ml of distilled
water from which 0.2ml was added to culture and
was kept for 20 minuites (Graph 1). Hypotonic
Solution: Hypotonic solution in compare to that of
cytoplasm of normal cell is used as a pre treatment
agent. Due to endosmosis, water from the
hypotonic solution entered to the cytoplasm of cell
through semipermeable plasma membrance,
resulting in swelling of the cells and subsequent
dispersion of the chromosomes. Potassium chloride
was used in a concentration of 0.600mg for the
above purpose (Graph 2).
GRAPH 1: THE CONCENTRATION OF 0.01 MICROGRAM
WITH 15-20 MINUTES EXPOSURE GIVES VERY GOOD
RESULT
GRAPH 2: THE BEST EFFECTS ARE OBSERVED IN THE
CONCENTRATION OF 600-625 MG% OF KCL FOR 40
MINUTES IN OUR STUDY.
Fixatives: For the preservation of the cell and their
contents in original form and to make them stable
during further processing, fixatives were used. In
the present study the reagents used as fixatives
were glacial acetic acid and absolute methyl
alcohol in the ratio of 1:3.
Staining: In above study conventional Giemsa
stain was used for the staining of slides. This
Giemsa stain was prepared from stock solution by
Naik et al., IJPSR, 2016; Vol. 7(1): 206-220. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 208
addition of 2ml of stock concentrate to 38 ml of
Phosphate buffer (pH -7.2). The prepared stain was
filtered using Whatman filter paper. This filtered
stain solution was added over air dried slide for
five minutes after which slides were rinsed with
distilled water twice and again kept air dried. The
stained slides were screened for wide spread
metaphase.
G. Banding: After dropping of cell suspension
over hot and moist slides, they were air dried and
kept in incubator at (37ºC) for 7days for
maturation. This matured slides after 7days
incubation were treated with 0.05 mg% of trypsin
(1:250) solution. Chemical denaturation of A=T
and C G bonds in between polynucleotide chains
occured at places as a reasult of treatment with
proteolytic enzyme (trypsin). On Giemsa staining
to this trypsin treated slides, alternate light and dark
bands of chromosomes were visible under
microscope and now the slide are called as banded
slide and the procedure is G. Banding.
Cytogenetic analysis:
The Bone Marrow was aspirated aseptically from
Posterior Superior illiac Spine of CML patient by
sterilized bone marrow niddle and a 10ml
disposable Syringe and 0.5 - 1 ml of the sample
was transferred immidiately to a culture bottle
containing 5 ml of preheparinized RPMI 1640
culture media. Then the bone marrow was
thoroughly mixed with the culture media by
shaking. This bone marrow in culture media was
then subjected for direct harvesting. To the culture
bottle containing bone marrow and heparinized
RPMI 1640 Culture media, colchicine was added in
a dose of 0.002 mg/ ml of bone marrow and mixed
thoroughly. It was then kept in incubator at 37ºC
for 20 minutes.
After 20 minutes of incubation the culture bottles
were removed and samples were transferred to
clean and sterilized centrifuse test tube. The test
tubes were marked by marker pencil and then
subjected for centrifugation in 900 rpm for 9
minutes. The supernantant was discarded with a
micropipette and the remaining cell pellet was
broken properly by fingers to make a homogenous
cell suspension. The hypotonic salt solution of 0.06
% KCL was added to the above homogenous cell
suspension and mixed thoroughly with the help of
micropipette. This hypotonic treated cell
suspension was kept in incubator at 37ºC for 40
minuites to swell up the cells. The swelled cells
after incubation was centrifused for 9 minutes at
900 rpm and supernatant fluid was removed. The
cell pellet thus obtained was broken slowly and
carefully to avoid rupture of fragile cell membrance
of swollen cells and resuspended in 0.5ml of same
fluid. To the above solution, chilled fixative (3
Parts of methyl alcohol, 1 part of glacial acetic
acid) was added slowly drop wise along the wall of
the inclined centrifuse test tube to the cell pellet
with a micropipette and kept for 10 min at room
temperature.
The chilled fixative treated cell suspension was
again mixed with 5 ml of fresh fixative and
centrifused for 9 minutes at 900 rpm. After
removing Supernatant fluid the remaining cell
pellet was broken properly to make a homogenous
cell suspension to which 0.5ml of fresh fixative
was added for resuspension and kept for 2hrs in
refrigerator. After 2hrs the resuspended cell
suspension was mixed with 5 ml of fresh fixative
and centrifused for 9 minutes in 900 rpm. The
Supernatant was discarded and the remaining cell
pellets were broken properly to get homogenous
cell suspension. 5 ml of fresh fixative was added to
the suspension and recentrifused after thorough
mixing. This above procedure was repeated for 3-4
times till a clear white cell pellet was obtained. 0.5
ml of fresh fixatives was added and mixed to obtain
a homogenous cell suspension. This clear cell
suspension was now ready for dropping.
Slide Preparation:
Air drying method for slide preparation was
followed for this study. New clean and grease free
slides were used for dropping. 2-3 drops of fixative
treated cell suspension were dropped on hot and
moist slide from the height of 20-24 inches. The
slides were kept in vertical position with slight
inclination during dropping and also kept in same
position after dropping. The dried slides were
coded properly by marker pencil.
Banding Protocol:
The coded slides were kept inside the incubator at
37ºC for 7days for maturation. The mature slides
Naik et al., IJPSR, 2016; Vol. 7(1): 206-220. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 209
were treated with trypsin solution (50mg trypsin in
100 ml distilled water) in a cuplin jar for 20
seconds. The trypsin digested slides were rinsed
with 0.9 % NaCl solution twice which were kept in
two cuplinjars. The slides were then washed with
distilled water in a cuplin jar and stained with 2 ml
of 4% Giemsa stain. The stain was poured over the
slide and kept for one minute after which 2 ml of
distilled water was added and a mixed thoroughly
and allowed for air drying. The stained air dried
slides were examined under light microscope for
Screening and selected for microphotography.
During taking microphotograph each slides were
screened for 15-20 well spreaded metaphases out of
which five of them were photographed. One out of
5 photographs having minimal over lapping was
karyotyped.
Karyotyping:
For this study, conventional ISCN-91 (International
System For Human Cytogenetic Nomenclature)
and Human chromosome (2ndedition) was reffered.
Karyotyping was done manually basing on the
following criteria. Lenth of the chromosome,
Position of the centromere, Presence of satelliate
bodies, banding pattern. The results were
confirmed by microscopic examination and
photographic observation. More importance was
given to direct microscopic observation, as the
photographs are less reliable than direct
observation.
RESULT: Chronic myeloid leukemia CML is a
clonal myeloproliferative disorder of the
pluripotent haematopoietic progenitor cells
characterized by excessive proliferation of marrow
granulocytes and presence of philadelphia
chromosome. Since the presence of philadelphia
chromosome is an important criteria for the
diagnosis of CML and also gives prognostic
information about its chemotherapy, CML patients
are usually subjected for cytogenetic analysis for
the detection of philadelphia chromosome. In the
present study 89 CML patients diagnosed clinically
and pathologically were analysed cytogenetically.
Out of 89 cases 54 were males and 35 were
females. The male to female ratio was 1.54:1. In
this stydy bone marrow samples were collected
from the patients with informed consent. In case of
minor or seriously ill patient, consent was taken
from the parents or from their guardian.
Cytogenetic analysis of all the patients were done
according to the standard protocol (ISCN-1991) 2.
The chromosomes were banded, karyotyped and
studied in detailed for structural and numerical
aberration.
The detection of philadelphia chromosome along
with presence of other chromosomal aberrations
were taken into account in the present study. In the
expression of the results, the patients having similar
findings were grouped and arranged in tables,
interpreted in ba1r charts and represented by pie
diagrams for comparision.
TABLE 1: PHILADELPHIA CHROMOSOME POSITIVITYIN
CML PATIENTS
Chromosomal Aberration
No of Cases
Percentage
Philadelphia Positive
72
81%
Philadelphia Negative
17
19%
Total
89
100%
This Table 1 shows the incidence of philadelphia
chromosome in clinically and pathologically
diagnosed CML patients. In the present study 89
cases of clinically and pathologically diagnosed
patients were analysed cytogenetically and
karyotyped. Out of 89 patients 72(81%) cases have
translocation t(9:22), i.e. philadelphia positive.
Among these 72 Ph positive cases, some are only
Ph Positive without any other chromosomal
aberration, while some others have additional
chromosomal aberrations along with the presence
of philadelphia chromosome. They may be either
structural or numerical aberrations. The remaining
17 (19%) patients are cytogenetically philadelphia
negative. Again among this group some patients
showed other structural and numerical aberrations,
while rest arekaryotypically normal.
TABLE 2: AGE SPECIFIC INCIDENCE AND MALE
FEMALE DISTRIBUTIONIN CML PATIENTS DIAGNOSED
PATHOLOGICALLY.
Age
Male
Female
Male To Female
Ratio
Total
0-20 yrs
2
2
1:1
4
21-40yrs
9
6
1.5:1
15
41-60yrs
25
16
1.56:1
41
>60yrs
18
11
1.63:1
29
Total
54
35
1.54:1
89
This Table 2 shows the Age and Sex distribution of
the pathologically diagnosed CML patients. In the
Naik et al., IJPSR, 2016; Vol. 7(1): 206-220. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 210
present study 54 males and 35 females are
analysed. The male to female ratio is 1.54:1. The
patients are within 2-70yrs of age. Median age is
47.5yrs. In the age group of 0-20yrs, there are
4(4.5%) patients. Out of them 2 are males and 2 are
females having the male to female ratio
1:1.15(17%) patients are within the age group of
21-40yrs. 9 males and 6 females with male to
female ratio of 1.5:1 are present in this group.
Maximum patients (46%) are within the age group
of 41-60yrs. With 25 males and 16 females this
group comprises 41 patients. Male to female ratio
is 1.56:1. The no. of patients having age more than
60 yrs are 29 (32.5%). There are 18 males and 11
females patients in this age group. The male to
female ratio is 1.63:1.
TABLE 3: CYTOGENETIC OBSERVATION VS AGE DISTRIBUTION
This Table 3 shows the distribution of different
chromosomal aberrations in different age groups.
There are 30(34%) patients having philadelphia
chromosome positive as the only karyotypic
anomaly. Among them 2 patients are in the age
group of 0-20 yrs, 7 patients are within 21-40yrs,
18 patients are within 40-60yrs and 3 patients have
age more than 60yrs.In the present study 9(10%)
patients are Karyotypically normal. They have
neither the philadelphia chromosome nor any other
chromosomal anomaly. In this group 2 patients are
within 21-40yrs, 5 patients are within 40-60yrs and
2 patients have age more than 60yrs.
Maximum number of patients have the philadelphia
chromosome positive along with other
chromosomal aberrations. Out of 42(47%) patients
of this group, 2 patients are below 20yrs, 5 patients
are within 21-40yrs, 15 patients are within 41-60yrs
and 20 patients have age more than 60yrs. Though
8 (9%) patients have ph -ve, they have some other
chromosomal aberrations. One patient of this group
has age within 21-40yrs, 3 patients have age within
40-60yrs and 4 patients have age more than 60yrs.
TABLE 4: CYTOGENETIC OBSERVATION VS MALE
FEMALE DISTRIBUTION
Chromosomal Aberration
Male
Female
Total
Ph +ve as only Karyotypic
anomaly
18
12
30
Ph -ve with normal karyotype
5
4
9
Ph +ve with other chromosomal anomaly
25
17
42
Ph -ve with other aberration
6
2
8
Total
54
35
89
This Table 4 shows the male female distribution in
different types of chromosomal aberrations of CML
patients. Out of 30(34%) patients showing
philadelphia chromosome as only Karyotypic
anomaly, 18 are males and 12 are females. Male to
female ratio is 1.5:1.
5 males and 4 females with male to female ratio of
1.25:1 are present in the group of CML patients
having normal Karyotype. Among the 42(47%)
patients having additional chromosomal aberrations
along with philadelphia chromosome, 25 are males
and 17 are females. The male to female ratio is
1.47:1.
8 patients are there in the group of CML patients
where philadelphia chromosome is absent but other
chromosomal aberration present. In this group there
are 6 males and 2 females with male to female ratio
3:1.
TABLE 5: CYTOGENETIC OBSERVATION VS MEAN &
MEDIAN AGE DISTRIBUTION
Chromosomal Aberration
Age
Range
Mean
Age
Median
Age
Ph +ve as only Karyotypic
anomaly
2-68
42.5
46
Ph -ve with normal karyotype
36-65
50.5
53
Ph +ve with other chromosomal anomaly
8-70
52.7
58.5
Ph -ve with other aberration
38-69
55
58
Total
2-70
50
54
The above Table 5 describes the mean & median
age distribution among the different types of
chromososmalaberration. In the present study 89
patients are undertaken within 2-70yrs of age.
Chromosomal Aberration
(Ph +ve as only)
0-20yrs
21-40yrs
40-60yrs
>60yrs
Total
Karyotypic anomaly
2
7
18
3
30
Ph -ve with normal
Karyotype
0
2
5
2
9
Ph +ve with other aberration
2
5
15
20
42
Ph -ve with other aberrations.
0
1
3
4
8
Naik et al., IJPSR, 2016; Vol. 7(1): 206-220. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 211
Mean age is 50yrs and median age is 54yrs. The
patients having philadelphia chromosome as only
karyotypic anomaly are within 2-68yrs of age
having mean age 42.5yrs and median age 46yrs.
The CML patients having normal karyotype are
within 36-65yrs having mean and median age
50.5yrs and 53yrs respectively. Other
chromosoamal aberrations present along with
philadelphia chromosome in a group of patients
within 8-70yrs having mean age 52.7yrs and
median age 58.5yrs.The Ph -ve CML patients
having other chromosomal aberrations were within
38-69yrs with mean and median age 55yrs and
58yrs respectively.
TABLE 6: CML PATIENTS HAVING PHILADELPHIA CHROMOSOME AS ONLY KARYOTYPIC ANOMALY.
Sl. No
Case
No.
Age
Sex
BM No.
Karyotype
No of abnormal
Karyotype per 20
metaphases
Remark
1
1
29
M
176/2006
t(9:22)46XY
15/20
75%
2
3
47
M
188/2006
t(9:22)46XY
16/20
80%
3
8
57
F
219/2006
t(9:22)46XX
17/20
85%
4
9
2
M
224/2006
t(9:22)46XY
13/20
65%
5
13
23
M
249/2007
t(9:22)46XY
14/20
70%
6
14
66
F
256/2007
t(9:22)46XX
18/20
90%
7
16
45
M
269/2007
t(9:22)46XY
16/20
80%
8
18
48
F
284/2007
t(9:22)46XX
16/20
80%
9
19
22
F
290/2007
t(9:22)46XX
14/20
70%
10
22
44
M
312/2007
t(9:22)46XY
15/20
75%
11
29
68
F
41/2007
t(9:22)46XX
17/20
85%
12
35
28
F
86/2007
t(9:22)46XX
15/20
75%
13
38
49
M
108/2007
t(9:22)46XY
15/20
75%
14
43
47
F
146/2007
t(9:22)46XX
14/20
70%
15
49
41
M
185/2007
t(9:22)46XY
13/20
65%
16
51
49
F
199/2007
t(9:22)46XX
14/20
70%
17
52
24
M
208/2007
t(9:22)46XY
13/20
65%
18
55
43
M
238/2007
t(9:22)46XY
15/20
75%
19
57
63
M
256/2007
t(9:22)46XY
17/20
85%
20
61
53
M
281/2007
t(9:22)46XY
16/20
80%
21
62
45
M
288/2008
t(9:22)46XY
15/20
75%
22
63
55
F
295/2008
t(9:22)46XX
14/20
70%
23
68
39
F
324/2008
t(9:22)46XX
14/20
70%
24
71
23
M
341/2008
t(9:22)46XY
13/20
65%
25
73
52
M
356/2008
t(9:22)46XY
15/20
75%
26
77
52
F
20/2008
t(9:22)46XX
16/20
80%
27
81
42
M
49/2008
t(9:22)46XY
16/20
80%
28
84
18
F
68/2008
t(9:22)46XX
13/20
65%
29
87
43
F
90/2008
t(9:22)46XX
14/20
70%
30
89
60
M
106/2008
t(9:22)46XY
16/20
80%
In the present study 30(34%) patients have
philadelphia chromosome as only Karyotypic
anomaly (Fig. 1 & 2). These patients are within 2-
68yrs of age and median age is 44.5yrs. 60%
patients are within 40-60yrs. Male to female ratio is
1.5:1. It is observed from the above Table 6 that
the number of abnormal karyotype per 20
metaphases is more in advance age.
TABLE 7: CML PATIENTS WITH NORMAL KARYOTYPE
Sl.
No
Case
No.
Age
Sex
BM No.
Karyotype
No of normal Karyotype per 20
metaphases.
Remark
1
7
54
M
212/2006
46 X Y
17/20
85%
2
17
65
M
275/2007
46 X Y
15/20
75%
3
41
40
M
132/2007
46 X Y
18/20
90%
4
48
56
M
179/2007
46 X Y
16/20
80%
5
54
48
F
226/2007
46 X X
17/20
85%
Naik et al., IJPSR, 2016; Vol. 7(1): 206-220. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 212
6
66
41
M
319/2008
46 X Y
17/20
85%
7
72
62
F
346/2008
46 X X
14/20
70%
8
78
36
F
28/2008
46 X X
18/20
90%
9
83
53
F
62/2008
46 X X
17/20
85%
Though philadelphia chromosome is chief criteria
for diagnosis of CML, it is not always found. In the
present study 9 CML patients are karyotypically
normal (Fig. 3 & 4).
FIG.3: G. BANDED KARYOTYPE OF BONE MARROW CELLS OF FEMALE CML PATIENT SHOWING NORMAL
KARYOTYPE 46XX.
FIG.4: KARYOTYPE OF BONE MARROW CELLS OF MALE CML PATIENT SHOWING NORMAL KARYOTYPE 46XY.
These patients are within 36-65 yrs of age. Median
age is 53yrs. Male to female ratio is 1.25:1. In the
above Table 7 it is observed that number of normal
Karyotype per 20 metaphases is more in relatively
younger age group.
TABLE 8: PHILADELPHIA POSITIVE PATIENTS WITH OTHER NUMERICAL ABERRATIONS.
Sl. No.
Case No.
Age
Sex
BM No
Karyotype
No of abnormal Karyotype per20
metaphases
Remark
1
2
42
F
182/2006
t(9:22)47XX+8
16/20
80%
2
5
21
M
201/2006
t(9:22)45XY-7
13/20
65%
3
12
61
F
241/2007
t(9:22)47XX+8
17/20
85%
4
15
56
F
262/2007
t(9:22)45XX-4
16/20
80%
5
20
68
M
296/2007
t(9:22)47XY+8
18/20
90%
6
23
58
F
322/2007
t(9:22)45XX-7
16/20
80%
7
26
67
F
19/2007
t(9:22)46XX+8
17/20
85%
8
28
24
F
34/2007
t(9:22)44XX-3
13/20
65%
9
30
58
M
49/2007
t(9:22)47XY+8
16/20
80%
10
32
64
M
66/2007
t(9:22)45XY-7
17/20
85%
11
33
50
M
72/2007
t(9:22)47XY+8
15/20
75%
12
36
51
F
94/2007
t(9:22)44XX-7
15/20
75%
13
39
64
M
114/2007
t(9:22)47XY+8
17/20
85%
Naik et al., IJPSR, 2016; Vol. 7(1): 206-220. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 213
14
44
30
F
151/2007
t(9:22)47XX+8
15/20
75%
15
46
12
M
167/2007
t(9:22)47XY+8
13/20
65%
16
50
67
M
192/2007
t(9:22)45XY-4
18/20
90%
17
53
63
F
216/2007
t(9:22)47XX+8
17/20
85%
18
56
64
F
244/2007
t(9:22)44XX-7
17/20
85%
19
60
59
M
273/2007
t(9:22)47XY+8
16/20
80%
20
65
66
F
307/2007
t(9:22)46XX-3
17/20
85%
21
69
46
F
329/2008
t(9:22)47XX+8
15/20
75%
22
74
65
F
362/2008
t(9:22)45XX-3
18/20
90%
23
75
46
M
3/2008
t(9:22)47XY+8
16/20
75%
24
79
44
F
36/2008
t(9:22)44XX-4
14/20
70%
25
82
66
M
356/2008
t(9:22)46XY+8
15/20
75%
26
85
69
M
74/2008
t(9:22)45XY-7
18/20
90%
27
88
30
M
96/2008
t(9:22)47XY+8
14/20
70%
Along with the philadelphia chromosome other
chromosomal aberrations are also found in CML
patients. Such aberration may be structural or
numerical. In the present study 27(30%) cases
show other numerical aberrations in addition to the
presence of philadelphia chromosome (Fig 5 and
6).
FIG.5: G. BANDED KARYOTYPE OF BONE MARROW CELLS OF FEMALE CML PATIENT SHOWING TRANSLOCATION
t(9:22) & TRISOMY 8. [t(9:22) 47XX + 8].
FIG.6: G. BANDED KARYOTYPE OF BONE MARROW CELLS OF MALE CML PATIENT SHOWING TRANSLOCATION
t(9:22) & DEL-3 [t(9:22) 44XY -3].
These patients are within 12-69 yrs of age. Median
age is 57yrs and mean age is 52.25yrs. Maximum
(81%) patients of this group are within 40-70yrs of
age. Male to female ratio is 0.93:1. The most
common numerical aberrations found in this group
is trisomy 8. Out of 15 cases having trisomy 8 in Ph
+ve clone, 9 are males and 6 are females. Male to
female ratio among them is 1.5:1.The next common
Naik et al., IJPSR, 2016; Vol. 7(1): 206-220. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 214
numerical aberration found in the above table is
del-7. Out of 6 cases of del-7 in Ph+ve group, 3 are
male and 3 are female having the male to female
ratio of 1:1.Besides the above numerical
aberrations in philadelphia positive patients, 3
cases have del-4 and 3 cases have del-3.
Out of 3 patients having del-4, only one patient is
male and 2 are females, while all cases of del-13
are females. So the male to female ratio has
reversed here with female predominance. The
number of abnormal Karyotype per 20 metaphages
is more towards late age.
TABLE 9: PHILADELPHIA POSITIVE CML PATIENTS WITH STRUCTURAL ABERRATIONS
The previous table shows the distribution of
structural chromosomal aberrations found in
philadelphia positive CML patients. Like the
numerical chromosomal aberration, there are also
different varieties of structural aberrations, found in
philadelphia positive CML patients (Fig.7 and 8).
FIG.7: G. BANDED KARYOTYPE OF BONE MARROW CELLS OF MALE CML PATIENT SHOWING TRANSLOCATION
T(9:22) & TRANSLOCATION t(1:7).[t(9:22)46XY t(1:7)].
FIG.8: KARYOTYPE OF BONE MARROW CELLS OF MALE CML PATIENT SHOWING TRANSLOCATION t(9:22) &
TRANSLOCATION t(15:17).[t(9:22)46XY t(15:17)].
Sl. No.
Case No.
Age
Sex
BM No
Karyotype
No of abnormal Karyotype per20
metaphases
Remark
1.
4
65
M
194/2006
t(9:22)46XY, t(15:17)
17/20
85%
2.
6
61
M
208/2006
t(9:22)46XY, t(2:6)
16/20
80%
3.
11
55
M
238/2007
t(9:22)46XY, t(15:17)
16/20
80%
4.
21
65
F
304/2007
t(9:22)46XX, t(15:17)
17/20
85%
5.
25
8
F
11/2007
t(9:22)46XX, t(15:17)
13/20
65%
6.
27
66
M
27/2007
t(9:22)46XY, t(6:7)
18/20
90%
7.
31
48
M
56/2007
t(9:22)46XY, t(15:17)
14/20
70%
8.
37
57
M
102/2007
t(9:22)46XY, t(1:7)
16/20
80%
9.
40
43
M
124/2007
t(9:22)46XY, t(15:17)
15/20
75%
10.
45
70
M
158/2007
t(9:22)46XY, t(2:6)
18/20
90%
11.
58
28
M
262/2007
t(9:22)46XY, t(6:7)
14/20
70%
12.
67
64
F
320/2008
t(9:22)46XX, t(15:17)
17/20
85%
13.
70
62
M
336/2008
t(9:22)46XY, t(1:7)
16/20
80%
14.
80
61
M
42/2008
t(9:22)46XY, t(15:17)
16/20
80%
15.
86
51
M
81/2008
t(9:22)46XY, t(6:7)
15/20
75%
Naik et al., IJPSR, 2016; Vol. 7(1): 206-220. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 215
In the present study 15(17%) cases show structural
chromosomal aberrations in addition to the
philadelphiachromosome. The most common
structural chromosomal aberrations found in Ph+ve
patients is t(15:17). Out of 8 patient having
t(15:17), there are 5 males and 3 females having
male to female ratio 1.6:1.These patients are within
8 - 70 yrs of age. The mean age is 53.6yrs and
median age is 59 yrs. Male to female ratio is
1.6:1.The rest of the patients showing additional
structural aberration are males. Among them 3
cases show t(6:7), 2 cases show t(2:6) and 2 cases
have t(1:7)along with philadelphiachromosome. In
the above table it is observed that structural
aberration are more seen in males and more number
of abnormal Karyotypes per 20 metaphases are
seen in older age in comparision to younger age.
TABLE 10: PHILADELPHIA NEGATIVE CML PATIENTS WITH NUMERICAL& STRUCTURAL ABERRATION.
Sl.
No.
Case
No.
Age
Sex
BM No
Karyotype
No of abnormal Karyotype
per20 metaphases
Remark
1
10
42
M
231/2007
t(2:6)46XY
15/20
75%
2
24
62
M
4/2007
t(15:17)46XY
17/20
85%
3
34
54
F
80/2007
47XX + 8
16/20
80%
4
42
67
M
139/2007
t(2:6)46XY
18/20
90%
5
47
63
M
172/2007
t(15:17)46XY
17/20
85%
6
59
69
F
268/2007
45XX – 7
18/20
90%
7
64
38
M
301/2008
t(2:6)46XY
14/20
70%
8
76
44
M
9/2008
47XY + 8
15/20
75%
There may be both structural and neumerical
aberration found in a single patient in addition to
philadelphia chromosome but in present study we
have not detected any such case. The structural and
numerical chromosomal aberrations are also found
in philadelphia negative CML patients (Fig.9 and
10).
FIG.9: KARYOTYPE OF BONE MARROW CELLS OF MALE CML PATIENT SHOWING TRANSLOCATION t(2:6).
[t(2:6)46XY)]
FIG.10: G. BANDED KARYOTYPE OF BONE MARROW CELLS OF FEMALE CML PATIENT SHOWING DEL-7.(44XX-7)
Naik et al., IJPSR, 2016; Vol. 7(1): 206-220. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 216
In the present study 8(9%) cases show such
abnormality having 6 males and 2 females. Male to
female ratio is 3:1. The patients are within 38-69
yrs of age. Mean age is 55yrs and median age is
62yrs. 5 cases show structural aberrations and 3
cases show neumericalaberrations.Thestructural
chromosomal aberrations seen in philadelphia -ve
clone are t(2:6) in 3 cases and t (15:17) in 2 cases.
The numerical aberrations found in philadelphia -
ve patients are trisomy 8(2 cases) and del-7
(1Case).From the above table it is observed that the
additional chromosomal aberration in Ph -ve clone
are found more in males.
DISCUSSION: Chronic myeloid leukemia is a
disease of clonal proliferation of
multipotenthaematopoietic stem cells characterized
by marrow hyperplasia of granulocytes, erythroid
precursors, megakaryocytes and connective tissue
forming cells. Association of philadelphia
chromosome with CML is well established. This is
the 1st consistent abnormality observed in human
cancer and is present in the bone marrow of 90-
95% cases of CML patients. After it’s 1st
identification in 1960, many researchers worked in
this field and found the same specific abnormality1.
Cytogenetic study play a dominant role in the
identification of philadelhia chromosome which is
diagnostic as well as prognostic indicator of CML.
In the present study 89 samples of bone marrow
from clinically diagnosed CML cases were
analysed. The conventional method (ISCN-1991)
was followed in this study. Out of 89 patients, 72
(81%) cases has 9:22 translocation and 17(19%)
cases were Ph -ve (Table - 1).
D. Costa GG et al. carried out their cytogenetic
study on 102 cases of CML 3. The samples were
analysed cytogenetically which showed 94%
patients having Ph chromosome positive.
In the study of ANGLO M et al.10 CML patients
were investigated 4. Cytogenetic study was carried
out in bone marrow samples using short term
culture. It was observed that 9 out of 10(90%)
patients have positive Philadelphiachromosome.
The cytogenetic analysis of 23 pathologically
diagnosed cases of CML were done by Jorge E
Corte et al 5. They observed that 17(73%) patients
were Ph +ve. Bone Marrow samples were collected
and analysed cytogenetically. 28(90%) cases shows
philadelphia positive. Similar to our observation,
16(84.2%) cases were Ph+ve in an analysis of 19
CML patient diagnosed pathologically 6. Nisha
Singh (2006) studied 16 CML patients diagnosed
pathologically 7.
In his conventional cytogenetic analysis Ph
chromosome was found in 13 (81%) cases and the
same (81%) Ph positivity was obtained in our
study. Our observation almost corroborates with
the data obtained by the above workers but there
are little variations, which may be due to a small
group subjected for analysis and geographical
variations.
In the present study age specific incidence and
male-female ratio of CML were observed (Table-
2).The maximum cases were observed in 41 to 60
yrs of age group. The male-female ratio was almost
equal in 0-20yrs of age group where it is maximum
i.e. 1.63 in the patients having age more than
60yrs.Similar age distribution in CML patients
were also seen in the study where 137 cases
between 15-68 yrs median (33yrs) were analysed 7.
Maximum patients i.e. 67(48%) were within 40-
60yrs of age but incontrast to our finding in age
distribution had undertaken a cytogenetic study on
34 pathologically diagnosed CML children having
age group between 5-11.5 yrs (median 8.2yrs) 9.
Male to female ratio was 1.2:1. So the male female
ratio of CML patients in 0-20yrs of age group is
almost similar to our finding even though the age
distribution in both studies are different. However
both age distribution and male female ratio in that
age group was similar to our observation with
another study where 19 CML patients within 60-
65yrs of age with male to female ratio 2:1 were
analysed 10. Late age distribution in CML patients
were also noticed in the study 11.
They analysed 30 diagnosed CML patients in the
age group of 26 - 57yrs out of which 14 (46%)
patients belonged to 5th and 6th decades. So our
observation tallies with some observers in age
distribution where with other observer in age
specific male female ratio. In few observers both
age distribution as well as male female ratio
Naik et al., IJPSR, 2016; Vol. 7(1): 206-220. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 217
showed similarity with the present study. The small
variation may be due to different geographical
distribution. One of the most important observation
of the present study is that, maximum 42(47%)
cases had Ph +ve with other chromosomal
aberrations and such patients had age more than
60yrs. (Table 3) So the patients having Ph
chromosomes with other aberrations have nearly
same or slightly higher percentage than the patients
having Ph chromosome as only anomaly. The same
thing was reported by S.I. Sonata et al 12.
They studied 67 cases of chronic myeloid leukemia
diagnosed pathologically. The patients were in the
age group of 20-68yrs. Out of 67 patients 28 had
Ph +ve as only karyotypic anomaly and 29 patients
had other chromosomal aberrations along with
philadelphia chromosome. However exactly same
percentage of distribution between only Ph positive
group and Ph chromosome with other anomaly
group occurred in the study 13.
In their study they analysed 32 CML patients
cytogenetically. 15 patients had Ph +ve as only
karyotypic anomaly where as 15 cases had Ph +ve
with other chromosomal aberrations. Again the
result appeared in the same manner as our study in
the work of Ahmed Aribi et al where 19 CML
cases within age group of 60-65yrs were studied 10.
Out of 13 Ph +ve cases 6 (32%) had Ph +ve as only
karyotypic anomaly and 7(37%) had Ph +ve with
other chromosomal aberrations. So there is striking
similarity in this regard between above workers and
our present studies. In the present work there are 54
males and 35 females suffering from CML, having
male predominance, the male to female ratio being
1.54:1. (Table 4).
36 patients with myeloid metaplasia of age group
between 13-65yrs were studied by J PM
Geraedtscytogenetically 14. The male to female
ratio was 1.47:1.The similar male predominancy in
CML was also noticed by the following workers
where male to female ratio ranges from 1.4:1 to
1.8:1. Anwar N. et al analysed 137 cases of CML
diagnosed pathologically. They were between 15-
68yrs (Median 53yrs) 8. The male-female ratio was
1.7:1. SettinA et al conducted a study on 63 cases
of CML having 40 male & 23 female patients 15.
The male to female ratio being 1.74:1. In an
another study of 21 patients between the age group
of 29-82 yrs(Median age 62yrs) were analysed by
Jorge Cortes et al andthe male to female ratio was
1.8:116.Michael W. et al analysed 32 patients of
CML. The patients were in the age group of 18-
80yrs, male to female ratio being 1.4:1 13. However
very high male predominancy in CML was also
reported by many authors where male to female
ratio was 4:1, 2.8:1 and 3:1 respectively 4, 6, 11.
They have analysed 10 patients, 19 patients, and 30
patients respectively. These variation may be due to
small group of patients taken into consideration in
their study. Another finding in the present study
was the median age of the patients being 54yrs.
(graph I & J, Table 5) which is corroborating with
following workers. The patients were in the age
group of 18-68yrs, median age being 54yrs. In an
another study 137 cases of CML diagnosed
pathologically 8. The cases were between 15-68yrs
(median 53yrs) which is very close to our
observation. Similar finding was obtained in an
another work where 69 diagnosed cases of CML
having BCR-ABL positive were studied by Claudia
Haferlach et al 17.
The patients were between 28-70yrs (median age
was 58yrs).From the report of another study, 32
patients of chronic phases of CML were
investigated 13. The patients were within 18-80yrs.
Median age 57yrs.With a close observation and
analysis of the findings of the above workers it is
concluded that CML patients present nearly similar
median ages with minimal variations as this is a
disease of 5th and 6th decade. CML patients having
philadelphia chromosome as only
karyotypicanomaly.
In the present study 30(34%) patients have
philadelphia chromosome as only
karyotypicanomaly (Table 6, Fig.1 and 2).
S.I. Sonata et al studied 67 cases of CML. Out of
them 28(42%) have philadelphia chromosome as
only karyotypic anomaly which is little higher than
that in our study 12. Again M.S. Anand et al studied
32 CML patients. Out of them 26 were in chronic
phase and 6 in blast crisis 19. 16(50%) had
philadelphia chromosome as only karyotypic
anomaly which is of still higher percentage. Radio
Naik et al., IJPSR, 2016; Vol. 7(1): 206-220. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 218
Oudat et al. studied 31 cases of CML. 20(69%)
cases had Ph +ve as only karyotypic anomaly 18.
Thus it shows extermely higher percentage and had
marked differenece from that in our study.
Supporting to the above workers R.K. Marwaha et
al had undertaken 34 pathologically diagnosed
CML children in which 18(52%) cases had Ph +ve
as only karyotypic anomaly9. Anwar N et al
analysed 137 CML patients diagnosed
pathologically where 73 (54%) cases had Ph +ve as
only karyotypic anomaly 8.
Here we found a marked difference in the
percentage of absolutely Ph+ve CML patients from
the above workers. The relatively low incidence
was observed in the only Ph +ve clone. This may
be due to new recurring chromosomal aberrations.
CML patients with normal karyotype:
In the present study 9(10%) patients had normal
karyotype. They had neither philadelphia
chromosome nor any other chromosomal aberration
(Table 7, Fig.3 and 4).Supporting to our finding, 3
(13%) patients were karyotypically normal in the
cytogenetic analysis where 23 pathologically
diagnosed cases of CML were analysed 16.
Similarly in the study of MS Anand et al, 26
patients of CML in chronic phase and 6 in blast
crisis were recruited. 2(8%) patients had normal
karyotype in their cytogenetic analysis which is
very close to our finding 19. In an another study 137
cases of CML diagnosed pathologically. 16 (12%)
patients were karyotypically normal 8. The
percentage of CML patients having normal
karyotype in our study almost tallies with the above
mentioned workers.
Ph +ve CML patients with other numerical
aberrations:
In the present study 27 (30%) cases showed other
numerical aberrations in addition to the presence of
philadelphia chromosome (Table 8, Fig.5 and 6).
Corroborating completely to this finding of our
study, S.I. Sonata et al studied 67 cases of chronic
myeloid leukemia diagnosed pathologically 12.
After karyotyping they found that 20(30%) cases
had other numerical changes in addition to
philadelphia chromosome in the form of
hyperploidy with frequent involvement of
chromosome +8, +17, +19, +21. The same
percentage of CML patients was also noticed in the
study where 26 patients of CML in chronic phase
and 6 in blast crisis (BC) were recruited 19. In their
study 8 (30%) patients demonstrated with trisomy 8
and del-13.R.K. Marwaha et al had undertaken a
cytogenetic study on 34 pathologically diagnosed
CML children 9.
They observed that 6 cases were with trisomy 8 and
4 cases had del-13 in addition to philadelphia
chromosome. So additional numerical aberration
was seen in 30% cases which is matching cent
percent to our study. Close corrobation was also
seen 20. They had conducted a study on 174 cases
of CML patients on different phases. 54(31%)
patients showed additional chromosomal anomaly
like del-3, trisomy 8 and monosomy-7.Similar
similarity was again observed in a study of 69
diagnosed cases of CML having BCR-ABL
positive17. Cytogenetic data of 30 CML patients
had additional chromosomal changes with trisomy
8 (10 cases), del-13 (6 cases), del-9 (4 cases) and
remaining 10 cases showed structural chromosomal
changes like t(1:7) 4 cases, t(13:15) 6 cases. So
20(29%) cases had additional numerical
aberrations.
Our finding corroborates 100% in this particular
aspect with some of the above observer and tallies
very closely to other mentioned workers.
Ph +ve CML patients with other structural
aberrations:
In the present study 15 (17%) patients have other
structural aberration in addition to Philadelphia
chromosome (Table 9, Fig.7 and 8). Similar result
obtained as the outcome of a study 12. They
examined 67 cases of CML diagnosed
pathologically. In their study 9 (14%) cases had
structural chromosomal aberrations like t(2:8) and
t(2:7) in addition to the philadelphia chromosome.
Like this, 69 diagnosed cases of CML, having
BCR-ABL positive, were studied byClaudia
Haferlach et al 17.
In their study 10(15%) cases showed structural
chromosomal aberrations like t(1:7) in 4 cases and
t(13:17) in 6 cases along with philadelphia
chromosome. In the same manner Susan Branford
et al studied 25 patients of CML. Karyotypic
Naik et al., IJPSR, 2016; Vol. 7(1): 206-220. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 219
analysis showed 23(92%) patients with
philadelphia chromosome, Out of them 8 had
additional numerical aberrations and 4 (16%) had
additional structural chromosomal aberrations
which is more closer to our observation 21.
However nearly similar percentage obtained in this
group of CML patients in the study 4. They
analysed 21 CML patients. In their study 18(80%)
cases showed Ph +ve. Out of them, 6 cases had
additional numerical chromosomal aberrations and
4(19%) had additional structural chromosomal
aberrations.
But the report given by Richabourg S et al tallies
perfectly to our finding. They carried out their
cytogenetic study on 41 CML patients with variant
philadelphia chromosome22. 38 (93%) cases
showed philadelphia positive and in 3 patients
other aberration like trisomy 8 (2 cases), del-13 (1
Case) were detected where as other 4 patients had
t(13:17) and 3 patients had t(1:7) as additional
structural chromosomal aberrations in Ph +ve
patients. So in 17% cases additional structural
aberration was present. From the comparison of the
reports and opinion of the above workers and that
of our study, it could be concluded that the rate of
occurrence of additional chromosomal aberation in
CML patients runs parallel to almost all workers
mentioned above, with little variations, which is
minimal and usual.
Philadelphia negative CML patients with other
chromosomal aberrations:
Another finding in the present study is that, 8 (9%)
cases are Philadelphia negative but with other
chromosomal aberrations.(Table 10, Fig.9 and 10).
Such statement also found in the report of S.I.
Sonata et al 12. They studied 67 cases of CML
diagnosed pathologically.
In their analysis 6(9%) patients in Ph negative
clone had other chromosomal aberrations as 4
numerical and 2 structural aberrations. Like the
carbon copy to this report, B. Anger et al also
claimed the same thing in their analysis where they
had examined 69 clinically and pathologically
diagnosed CML patients 23. Out of 69 cases
57(83%) showed philadelphia negative. Among
them 6 (9%) had other chromosomal aberrations.
So both the above workers had cent percent
similarity with our findings. However though not
cent percent, very closely related result was
observed in the study of Nisha Singh who studied
16 CML patients diagnosed pathologically 7. He
found that 13 (81%) cases were Ph +ve, the
remaining 3 were Ph -ve. In Ph -ve group, 1 had
structural aberration, 1 had numerical aberration
and 1 had normal karyotype. So the percentage of
Ph -ve CML patients with other chromosomal
aberation was 12.5%. They observed that in 7
patients, philadelphia chromosome was absent.
Among them 2 (3%) patients had trisomy 8 and 3
(4%) patients had t(13;15) and 2 were
karyotypically normal.
Ahmed Aribi et al also reported the similar thing
from their analysis which tallies with above
workers 10. They had studied 19 CML patients and
found that 13(69%) cases had philadelphia
chromosome positive. In philadelphia negative
group 2(11%) patients had t(13:15). Even though
philadelphia chromosome is very much popular and
essential for the diagnosis of CML, a small group
of stamped CML patients are devoid of it. Instead
they may possess other structural and numerical
anomaly which were proved from the reports of the
above workers and corroborating to them, similar
finding were also obtained in our present study.
Finally it is observed that many of our findings are
corroborating to a number of previous workers, at
places they are matching exactly or at places with
little variation. However in findings like absolutely
Ph +ve CML cases we found marked variation
which may be due to emergence of newly occurring
chromosomal aberation in addition to philadelphia
chromosome.
CONCLUSION: The marked variation was
observed in the percentage of only Ph +ve group
and this may be due to emergence of additional
chromosomal aberration in Ph+ve CML patients.
We found trisomy 8 as most common additional
numerical chromosomal aberration which is also
reported by many of the previous observer. But the
most common additional structural chromosomal
aberration found here was t(15:17), which is
relatively rare in available literature. To establish
this as a newly recurring additional chromosomal
Naik et al., IJPSR, 2016; Vol. 7(1): 206-220. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 220
aberration in CML patients, large database analysis
is required.
ACKNOWLEDGEMENT: The authors would
like to thank Prof. Manoj Ranjan Nayak, president
and Er. Gopabandhu Kar, S‘O’A University for
extended facilities. Dr. Sitansu Ku. Panda, Asso.
Prof, Dept of Anatomy wrote the manuscript and
drafts it. Miss Subhrajita Rout, Tutor in
Microbiology, IMS & SUM Hospital helped for
writing the manuscript and draft it.
REFERENCES:
1. Nowell PC, Hungerford DA. A minute chromosome in human
chronic granulocytic leukemia. Science 1960; 132:1197.
2. ISCN (1995) Guidelines for Cancer Cytogenetics, Supplement to:
An International System for Human Cytogenetic Nomenclature
(ed. by F. Mitelman). S. Karger, Basel.
3. D’Costa GG, Siddiqui HM, Pradhan RM, Gupta SS: A ten year
incidence study of 242 cases. Pattern of leukemiayear 1989;
35(4):191-195.
4. Angelo MC, Frassioni F, Pollicardo N, Pungolino E, Ferrero R,
Vasallo F, Soracco M, Giordano D, Figari O, Benvenato F,
Florio G, Carlier P and Valvoness M: Philadelphia chromosome -
ve peripheral blood stemcells can be mobilised in early phase of
recovery after a mylosuppressive chemotherapy in philadelphia
chromosome +ve CML : BJ H1994;89:535-538.
5. 5.Cortes JE, Talpaz M, Beran M, Sasan M, Brien O, Mory B,
Stass RS and Hagop MK: Philadelphia chromosome -ve chronic
myelogenous Leukemia with rearrangement of the Break point
cluster region. CANCER1994; 75:464-469.
6. Jha CB, Bhatta NK, Karki P (): Cytogenetic study of philadelphia
chromosome in CML patients. IJPD2005;2(5) 2005-11 - 2005-12
7. Singh N: Association of philadelphia chromosome in CML.
Molecular Cytogenetics 2013; 6:59doi:10.1186/1755-8166-6-59
8. Anwar NM, Pamela, Jeffrey Z and Charles AS: effect
imatinibmesylate patients with philadelphia chromosome +ve
chronic myeoid leukemia with secondary chromosomal
aberration. CCR2003; 9: 1333-1337.
9. Marwaha RK, Bansal D, Trewhan A, Varma N: Profile and
survival patterns in children with Adult-variety chronic myeloid
leukemia, treated with conventional chemotherapy. Ped Hem Onc
2003; 20:505-15.
10. Aribi A, Borthahar G, Ravandi F, Shan J, Davission J, Cortes J,
Kantarjian H: Activity of Decitabine, a Hypomethylating Agent,
in chronic myelomonocytic leukemia. CANCER2007; 109:713-
716.
11. Oki Y, Hagop KJ, Gharibyan V, Jones D, Brien SO, Varstovsek
S, CortesJ, Morris GM, Guillermo K, Garciamanero H, Issa JP:
Phase II study of Low-Dose Decitabine in combination with
Acelerated or Myeloid Blastic phase of Chronic Myelogenous
Leukemia. CANCER2007; 109:899-905.
12. Sonata SI, Sandberg AA: Chromosome and causation of Human
Cancer and Leukemia. CANCER1978; 41:153-173.
13. Michael WN, Inger D, Cortes J, Paquette R, Park B, Hochhaces
A, Baccarani M, Stone R, Fischor T, Hagop K, Niederwiesar D,
Passcrini CG, Charlene S, Gathmann I, Goldman JM, Doulas S,
Brian Druker J, Guilhot F (): The prognosis for patients with
chronic myeloid leukemia who have clonal cytogenetic
Abnormalities in philadelphia chromosome -ve cells.
CANCER2007; 110: 1509-1581.
14. Geralldts JPM: An identical translocation between chromosome 1
and 7 in myeloid metaplasia. BJM1980; 44: 569-575.
15. Settin A, Haggar MA, Dasoky T, Baz AR, Desrakzik AN, Fouda
M, Arefs MA, Tonbary Y: Diagnostic cytogenetic markers in
CML. Med PedOnc2007;79(3): 255-263.
16. Cortes J, Alfonso QC, Guillenmo GM, Brien SO, Jones D, Faderl
S, Teresa E, Giles F, Deborah T, Hagop K: Phase-1 study of
Tipifarnib in combination with Imatinib for patients with Chronic
Myelogenous Leukemia in chronic phase after Imatinib failure.
CANCER2007, 110: 2000-2005.
17. Outdat R, Khan Z and Glassman A: Detection of trisomy 8 in
philadelphia chromosomal cytogenetic and interphase Florescence
In Situ Hybridisation Techniques and it’s relation to C-myc
Involvement. Annuals of clinical & laboratory Science 2001;
31:68-74.
18. Anand MS, Verma N, Verma S and Kaul D: Cytogenetic &
molecular analyses in adult chronic myelogenousleukaemia
patients in north India. Indian J Med Res. 2012 Jan; 135(1): 42–
48.
19. Haferlach T, Kenn W and Schoch W: Additional clonal
abnormality in phladelphia positive ALL and CML demonstrate a
different cytogenetic pattern at diagnosis and follow different
pathway at prograssion. Cancer Genetics and Cytogenetics2005;
157(1): 53-61.
20. Branford S, Hughes T, Mislner A, Koelmeyer R, Schwarer A,
Arther C, Shie RF, Moreton S, Lynch K, Taylor K: Efficacy and
safety of imatinib in patients with chronic myeloid leukemia and
complete or Near-Complete cytogenetic Response to Interferom.
CANCER 2006; 110: 801-806.
21. Richabourg M, Eclacha V, Perotc G, Portnoi MF, Akker VD,
Terre J, Maareck O, Vigaie F, Lat IL, Andrieux J. Cormn S and
Roche LC: Patterns of genesis of variant translocation in chronic
myeloid leukemia are not related with ABL-1 or BCR deletion
status or responce to Fmatinib therapy. BJH 2008; 15 (1), 126-
134.
22. Anger B, Carbonels F, Branger I, Heinze B, Guten W, Sohn, ET
and Heimpel H: Blast crisis of philadelphia chromosome positive
chronic myelocytic leukemia (CML). Annals of Haematology
2004; 57:724-735.
All © 2013 are reserved by International Journal of Pharmaceutical Sciences and Research. This Journal licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License.
This article can be downloaded to ANDROID OS based mobile. Scan QR Code using Code/Bar Scanner from your mobile. (Scanners are available on Google
Playstore)
How to cite this article:
Naik MK, Mishra DN, Rout S, Panda P, Panda SK, Das SR, Mishra T, Mohapatra C: Ph iladelphia Chromosome in
Pretherapeutic Cases of Chron ic Myelogenous Leukemia. Int J Pharm Sci Res 2016; 7(1): 206-20.doi: 10.13040/IJPSR.0975-
8232.7(1).206-20.
