research done by dr. jackleen raafat awadallah hanna phd of medical biochemistry researcher at...

Research done by Research done by

Dr. Jackleen Raafat Awadallah HannaPhd of Medical Biochemistry

Researcher at Medical Biochemistry DepartmentNational Research Center

Egypt

Under the Supervision of

Prof. Dr. Maged A. Barakat Prof. Dr. Zakaria A. El-Khayat Professor of Biochemistry Professor of Biochemistry Faculty of Pharmacy Medical Biochemistry Department Cairo University National Research Center

Dr. Manal F. IsmailAss. Professor of Biochemistry

Faculty of Pharmacy Cairo University

Expression of CD95 in Acute Lymphocytic Leukemia (ALL) in Egyptian Children before and

after Treatment

Leukemia

It’s is the term used for cancer that affects the blood cells.

Marrow-based neoplasm composed predominantly of minimally or partially differentiated lymphoid precursors.

The malignant disorder resulting from the clonal proliferation of lymphoid precursors with arrested maturation is known as

It is the most common type of Leukemia found in children that's why it is commonly called childhood leukemia.

Apoptosis

(Programmed cell death)

Controlled cellular self-destruction.

Regulation of normal cell growth is balanced between cell proliferation, cell differentiation and apoptosis, a disruption of this balance is thought to be an important event leading to carcinogenesis. Apoptosis induction is the target of most chemotheropeutic drugs.

Regulation of apoptosis

Pro- apoptotic protein.

CD95

(Fas, Apo-1)

Cross linking of Fas by its ligand (Fas L)

Apoptosis

Bcl-2

It is located within the mitochondrial membrane, endoplasmic reticulum and nuclear envelope.

Bcl-2 family of proteins :- Bax , Bcl-2

Treatment protocol for childhood ALL

The induction phase (6-7 weeks)

The consolidation phase(8 weeks)

HD-MTX , 6-MP

DEX, VCR, DOX, 6-MP, ASP, Ith, MTX, Cyclo, Ara-C.

VCR, DOX, Prednisone, Ara C,6-MP,Cyclo, ASP Ith (MTX, Ara C, hydrocortisone)The maintenance phase

(120 weeks)

Total XV (Pui et al., 2004)

Zinc Copper

Zinc is found in almost every cell of the body.

It forms complexes with metalloenzymes with oxidase activity. It stimulates the

activity of approximately 100 enzymes.

Nutritional status of children with ALL

Aim of the workIdentify the changes occurring in newly diagnosed ALL children regarding the expression of CD95

receptor and blood level of Bcl-2 as markers of apoptosis as well as serum zinc and copper levels prior to and during the maintenance phase of the specified treatment protocol.

Subjects

Group 1 Group 3Group 2

Ten normal healthy children

Twenty five pediatric patients newly diagnosed

of acute lymphocytic

leukemia

Twenty treated patients

Routine laboratory investigations

Complete blood count

(CBC)

Hemoglobin concentration

Bone marrow

blast count

Total leukocyte count

Platelet count

Sampling

Determination of CD95 % using

Flowcytometer Determination of Bcl-2

Concentration using ELISA

Blood

Determination of zinc and copper

levels using

atomic absorption spectrophotometer

Serum

Hemoglobin concentration in groups 1,2 and 3.

a

a,b

0

2

4

6

8

10

12

14

Hem

og

lob

in (

gm

%)

Group1

Group2

Group3

Platelet count (Plts) in groups 1,2 and 3.

aa,b

0

100

200

300P

lts.*

(10)

3 (c

u.m

m.)

Group1

Group2

Group3

Bone marrow blast count (B.M.blast) in groups 1,2 and 3.

aa,b

0

20

40

60

80

100

B.M

.bla

st %

Group1

Group2

Group3

Total Leukocytic count (TLC) in groups 1,2 and 3.

a

b0

50

100

150

200

250

TL

C *

(10)3

(cu

.mm

.)

Group1

Group2

Group3

CD95 % in groups 1,2 and 3.

a

a,b

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7C

D95 % Group1

Group2

Group3

Bcl-2 concentration in groups 1,2 and 3.

a

a,b

0

100

200

300

400

500

600

700

800

Bcl-

2 (

pg

/ml)

Group1

Group2

Group3

Serum Cu levels in groups 1,2 and 3.

a a

0

100

200

300

400

Cu (u

g/dl

)

Group1

Group2

Group3

Serum Zn levels in groups 1,2 and 3.

a

b

0

50

100

150

Zn (u

g/dl

) Group1

Group2

Group3

Cu/Zn in groups 1,2 and 3.

a

a,b

0

1

2

3

4

5

6

Cu

/Zn Group1

Group2

Group3

0.00 200.00 400.00 600.00 800.00 1000.00

Bcl2

0.00

0.20

0.40

0.60

0.80

1.00

CD

95

R Sq Linear = 0.618

40.00 60.00 80.00 100.00 120.00

Zn

100.00

150.00

200.00

250.00

300.00

350.00

400.00

Cu

R Sq Linear = 0.514

The current study demonstrated that both CD95 % and Bcl-2 protein are useful diagnostic markers that may help not only in diagnosis but also in follow up of ALL cases.

Long-term studies with large number of cases are still needed.

Avoiding chronic zinc deficiency should be taken into consideration in the prevention of childhood leukemia.