5 &amp amp; ) 82BCL D ' 6 E 01*% 82 1

/ /91 / !"#$ /99

99

1391

: 6 / 2 / 91

: 13 / 4 / 91

: 20 / 4 / 91

! "

#

$

% BCL1

1 :

! 2 3

*

1 .

!"

! #$

%"

&

' ()

'

2 . +"

+ +,

# - .+./) + +, 0 #+1 +,

%"

' () ' () '

3 . +"

!"

! #$

%"

&

' () '

* 4 54 6 : 8

E-mail: ryaraee@yahoo.com

$& '

&

(&

: !

!

"

# $

!

# !

% & ' ( ) *+

* & ' ,

-

! .*

# / 01 2 ! 3* 2 4

5

# & )

* 6$

7 0 5 62 8 $ 4 '

*& % 0 9 '

# 8 $ !

&

' - : '

*& % 62 , ' ! -' % 5 ;

# 5 ' &

5 ;

<=

6$

>*$

&

8$

"

!?

.*

, ! 6@ A' 0

$

6 4'

B 2

# ! & ) *+

, $

# @ 7) 5 ;

;! !

5 ;

6 4'

$ B!

( $ 5

& ) ! .

# : 1 & )

82BCL1 1 !

*%

5

# @ 7) ! 5 :) D ' 6 E 01*%

82 1 !

*&*%

$

1 !

!

(F '

6$

*&*% ! ! . B!

# ! ,

G H

#

I =' $ , ! 5

# 5 ' &

5 ; 5 :)

D '

6 4'

B!

,

! $ 1

J 6 BCL1 ( '

$

*%

.

*K

6 2

! L 1 !*&!MTT

# /

# E& ' IC50 I =' 67 :'

*%

.

*K 5

# 6$

D M&

# ' 6& /

* !

* ' $ , * 2 N& ! 1* 8' $

O '

P& Q 6$

*' .

* % : N &

( 8& MTT

! 62 G H 5

J / '

$ ' ,

B!

2

-$ S TE $ 1

! BCL1 G H

2

/ '

J $ ' &

$

%?/

( ' B!

2

>

! ( 8&

'

* .

# 5 ' &

5 ; 0 G H

# 20 10

J / '

$ '

! X $ 9 ' ' '

*& %

"

G H

# 2 ! 5 B!

2 &!* Y $ 5

# 5 ;

;! !

!*&

*&

; 62 G H

# 2 ! 5

J / '

$ ' 5

# 5 ;

*@$

! 72 B!

2 @'

# !

( 8&

*& ! .

*K 5

# [ '

B ! G H

# I ='

! 1* 8' 1*%

$

O '

P& Q $

*K

6 2

6$

1*' ( G H

#

!

\ 'MTT $ .

+ % , : 6$

' G&

* , 8 $ B!

2 , !

! $ 5

# & ) ! 5

#

@ 7) 6$

]*%

6$

( '

@Q 0

! 6 $!

.

- ,.

& /0 : 0 0

)BCL1 1 & ) J (0 0,

# 5 :)

D '

/ /91 / !"#$ /99

&

6 9 : + , &

: $ $ , ' ") +;

, ' $

<

0 #+1 6 =1 & 1 &

4&

. ': ) >

' + ? @ : $ $ : +

& +$ : ! A ' B

. , : $ $ 9+

6 9

&

& 1 B +C +D

" % =+EB 0 +, F

) 1 ( I

6 9 B &

, J+4.)

?, > +D 6

A . 6 9

=+EB &

K ) L % &

M 6 9 ,

N> J+4.)

» ( J4+ ! )

«

. 9 Q 9 J+4.) 1 6

&

6 9 Q =+EB &

9 ?!+ R B : B

L M )

2 .(

+ )

F " ,

6 9 B &

) , 4, , S

N>

A 9 J+4.) ' .

"+, , &

T+

,

RNA

L M F DNA

&

<

6 9 F , ,

&

U 4& J+4.) 6 ; )

? / +4, A )

, + V

"

<

. : $ $ F+ W & & F )

3 .(

E X ,

: 9 +Y!) N9 @ &

"+,

J(

A+ C $ F . ,

&

،

X ) J 4+ : ,

J+

Z +Y!) .

9

6 9 [ $ J( 1

J 4+ &

4& #+

. A+ C [ $ B &

&

A/)

> : + J+4.) +

,

\]^_

» 6 9 S 4

1« + .

1 : =, '

[ $

&

A+ C =) ، &

&

,

&

> , : )

4 .(

F+ 4 !

a , A B b

I 6 9 ?!+

) 5 .(

: F+ 4 !

% , T+ ) A & + ! !

L C ' ? d

C .

4& , ) !+ b ? d & + ! !

1. Clonal expansion

?_

6 , ) !+ =+EB !9 d , ? ) R B : &

9 Q A1 "+$ N> ) ' +

: 1

G2-M

V .

, X ) , J4+ !

V F+ 4 ! F ،

,

e V 0 ' d

: + 9; ,

6 9

&

+Y!) 6 ; ,

f &

, V

&

6 9 , ' ;

; =+EB a &

A; 6

+Y!) 6 ; =+EB g @

,

& +Vh) F

,

Z ' #+ i : B 9 9 +

4 . : $ $ j >

.

= ^ F f &

V ,

, F+ 4 ! !9, A4+ k B

, = ^ + h)

f & & V

A 9 J+4.) +

. :

d 4 ) V l 0+

C , A+

&

A V

' C , A+

Y!) 6 ; +

6 9

@ $ %

A J 4+

L 9=

> , A

.

#

F+ 4 ! a )

' @ A A

(

+() A" m+/ : C ,

RPMI1640

Ai9D

&

20 10 5 2 1 5 / 0 + 9+ , L ! +

: J& 1 .

6 9 S 6 /B &

BALB/c

, UE ' d

6 9 g @ =+EB &

, a , .

A '

6 9 : & , +Y!) 6 ; 6 9 S 6 /B &

F+ ! p / )A

(ConA

#1 F " : =, .

) ?+) , S ? q $ W+> : C ,

X S 6 /B ? m ,

0 ?

m+/ / ? $

RPMI 1640(Gibco)

?.

. C , 6 /B ' "1 , ? q $ :

6 9 , 6 /B &

' +4 r - _

A" m+/

&

. 6 9 X &

1 , ' !

15

?.

6 9 +C &

13 X

%+

1500 .+>

)

500 × g

( - , 10

L M .+>

. +C : =, U

@ 6 9 ,

+C &

1 ,

#+

6 E9 # > &

#1 .

: #+ 1 ,

/ /91 / !"#$ /99

q ) 206 / 0 ) % : M; –

^. g (

L + 9

83 / 0 _ , AE4 1 , 10 ) M;

(

, . 6 9 - , 6 /B &

2 .+>

> #+ 1 , l = ) 1

6 E9

# > &

#+

. ,

#+ : + 9X 6 9

&

T9D

4& A+ C , 9_ 1],

F+ X L

(FBS¹)

#1 .

6 E9 fe; , +C &

#+ # >

13 X

1500

.+>

) 6 = ( - , 10 L M .+>

. 9;

, " 4 m+/

T> = RPMI

' #+ +C

6 9 L M &

. 6 9 ,

> , &

m+/

, A"

10 _ #1 FBS

,.

_ ,

6 9

&

X v : 6 /B : :

6 9 #+

9, ' r ) , C w ! !+ , & "

w ! !+ : , 5 Lx , 9 S L M A1 :

! &

6 9 _ &

"+ & : x ,

98 , _ .

: A+9$

A"

6 9 96 ,

i A"

6 9

&

C . a , , A ' 6 9 6 ;

+Y!) ,

&

: 6 9

&

6 /B S

p /

F+ !

(A) ConA

#1 .

z , 6 9

&

6 /B

&

N9 @

? 6

6 ,

p / F+ !

(A) ConA

, Ai9D 25

L !+

,

9+

+

&

) Ai9D

&

20 10 5 2 1

5 / 0

L !+

, 9+

+ ( A (

&

&

, p / F+ !

(A) ConA

, .

&

0& Q

=) 105

× 2 6 9 ,

&

Ai9D

6 5 0& Q A"

. , ' :

&

24 48

72 Ad A+9$

X

? ) , ! , 5 _

>CO2

.

9

$ + 4 9 0 , : BCL1

' ) (C551

A" m+/

1. Fetal Bovine Serum

RPMI1640

)

Gibco

( 10

_ L

FBS

)

Gibco

( A"

. 6 9 A4) L M ,

&

:

X A" 0 ]1

S : =, )

' r ) , &

9, ( =) 10⁴

× 2 , 6 9 6 9 A" A+9$ 0& Q &

96 ' + , ! T 0 : =, ?.

37 X ?; m+/ } =)

Ai9D ' #1 N9 @ &

) 5 2 1 5 / 0

, L !+

9+

+ ( , "

F+"+$

L M .

! " MTT

=, : +) ' : A e )

24 48 72

Ad (

~ ) , ! : A+9$

& :

0& Q & , ? m 20

6 9/ + !+

MTT

) Ai9D 5 _ 6

F+

( #1 ,

- , ) , ! ? 4

> Ad A1

) ( . A+9$

F : )

% A+9$

,

+C :

+() . +C 27

X ,

- 10 .+>

2000 L M .+>

A1 . =,

0& Q ? m+/ +C : +9@) A> , &

, B 4

6 C , &

> , 0& Q v

, . 0& Q , + 6 $ $ # &

)

0.04N

+

0 9

(

6 4 ) 1 b

?; &

<

A ( ~ 6 B geX 492

,

# x % . –

geX ' #+

T

, i , A ) +; A+ =1 geX F+% + F 1

6 ) > 1 (

, ' d 100 _

E / +4+4 ) .

/ /91 / !"#$ /99

A B

C D

1"

1 . / $ 4 BCL1

5 6/7

- 5 8/ 9

48

<

. / ET/AO

=%

> ?

$& @ A% /B C

&%&D .

9 6

, ' #1 ' , BCL1

#, .x40

6 9 , #E q 91 , : &

B

& " F T D .

hV B

:

Ai9D 5 9+ , L !+

+

#, , .x40

+) $ $ ( 9 F # > q 91 , 0

, T D B .

9 C

hV :

Ai9D 2 9+ , L !+

+

#, , .x20

' &

& " B 6 9 =)

, : &

#E q 91

: "+, ; ) 6 9

, 0+) $ $ &

# > q 91 , <A

A4+ T D ? B .

9 D

hV : Ai9D 1 9+ , L !+

+

#, , .x40

6 9 q 91 , : &

A F T D A+= X #E .

$ $% &$ ' ' " ( "

6 9 &

Ai9D : T@ &

&

6 ) p / ' , , (

+9$

A" A A"

. B : =, 48

6 9 Ad &

,

PBS¹

" 4 L +) v T+ ) ,

, / v • F

#+

,

q 91 w ! !+

OLYMPUS

6

BX51

& "

, i 6 9 =) E4 F++=) &

M S : 0+) $ $ L

. J& • F

6 9 , 6 9 , J& &

€ C : &

" v #E q 91

? M+ &

'

) (

v F , F , , A DNA

, v F ( )

: +D : : 6 9 ' )

•+@") )

6 ( , , L +) .

, 6 ‚) : =

DNA

6 9 ++ƒ) &

&

"D ? /

) 6 9 0+) $ $ &

( " # > q 91

) 7 ( v : C , .

#+

ET/AO²

' )

1. Phosphate buffered saline

2. Ethidium bromide / Acridine orange

/ /91 / !"#$ /99

„ 1 ' + 6 9 : : $ $ : &

& " q 91 w ! !+

<

S : =,

_ E / q!d

&

#+

, : 1 ‚) - _

@, 1 A+9$ N9 @ &

.

? E F

, +4+4 ) _ S , geX K

T-test ANOVA

+4C) . • , N9 @ &

' : : C

ANOVA

L M A1

p<0.05

= , i : ' ,

1 i

.

B $ $% & * &+

&

=) 200 : 6 9 6 9

&

v

#+

,

ET/AO

,

1 ‚) B

:

S q 91 w ! !+

6 9 _ &

0+) $ $ 9 &

hV A e : =, :

48 , Ad A

, ' ' #+ •@"

Ai9D #1 ,

-

#1

< , - _ F ,

6 9 . F &

9 6 )

' ,

( 4 _ 9 h

: V Ai9D &

5 / 0

،

1 2، 5 9+ , L !+

, + T+) ) 18.5 24، 51،

52 , _ ) 6 X 1 .(

A+9$ : 6 9 ; &

N9 @ m &

) :

6 9 X (

q!d

&

, #+

‚) B 1

1 )

‚) 3

1 .(

' &

B

6 9 AE4 0+) $ $ A+=b &

…;

Ai9D , )x , &

A #1

.

)h , F+ 4 ! +V +; A+ =1

6 9 ) &

BCL1

' : 24

48، 72 Ad :

: C , 0& Q geX

Ai9D , &

N9 @ 6 9 ) +; A+ =1 _ ،

&

J ) 1

.(

' &

B 1

& "

Ai9D 5 9+ , L !+

+

0+4 ) V ' : &

= 6 9 ,

&

' " BCL1

<

; Ai9D 2 , L !+

9+

+ +) : =, 48

Ad

0+4 ) V

Ai9D A

&

1 9+ , L !+

+

V

= 0+4 ) ' "

. J& 4 . ' :

V , 4 . _ ?! , 6

' :

e

?, >1D

A & "

.

,

* - . '/.

& 0 1 23

"

24 48، 72 6

: C , geX

0& Q N9 @ Ai9D , &

6 9 ) +; A+ =1 _ ، 6 &

6 /B ' :

24 48 ، 72 Ad V T> =

Ai9D J F+ 4 ! N9 @ &

) 2

.(

' &

B

2

& "

Ai9D &

20

10 9+ , L !+

+ 24 48 ، 72 Ad

Z †d ,

= + 6 9

6 &

) 6 ;

A;

( 6 /B .

Ai9D

&

: 10 V

4 ) 0+

=

24 ' " Ad

: =, ' : A e , 48

72 e V Ad

6 9 ,

&

& " A; 6 ;

' "

, 6 9 A; 6 ; &

6 /B

' : Ai9D , 4, 0+4 ) V .

)h , F+ 4 ! +V ) +; A+ =1

6 9 6 ; &

' : 6 /B +Y!) 24

48، 72 Ad

6 9 ) +; A+ =1 _ 6 /B +Y!) 6 ; &

24 48، 72 Ai9D V T> = Ad N9 @ &

' " F+ 4 ! Ai9D &

: &

V

= 0+4 ) 6 9 F ,

&

.

+4 ) V

Ai9D , 4, 0 6 9

6 ; &

& " +Y!) Ai9D - _ F ,

)x , &

Z d , "+, +

† ) 3

.(

9 ) +; _ 4 .

) 6 /B

A; 6 ;

9 +Y!) 6 ; 6 /B ،

(BCL1

4 ' "

F+ 4 !

Ai9D

&

, ' 4!

9 , T+) ) 6 /B،

+Y!) 6 ;

A; 6 ;

V F "+,

A 0+4 ) .

/ /91 / !"#$ /99

A

B

C D

1"

2 . / F 4

5 6/7

- 5 8/ 9

48

<

.

A

B

C D

1"

3 . /

! " F 4

5 6/7

- 5 8/ 9

48

<

. /

ET/AO

=%

$& @ A% /B C > ?

&%&D .

#, , ' #1 ' , 6 A

. 40x

B

hV : Ai9D 20 9+ , L !+

+

#, , 40x

. 6 9 # > q 91 , 0+) $ $ &

, F T D B

. 9 C

hV : Ai9D 10 9+ , L !+

+

#, ,

20x

. 9 D

hV : Ai9D 5 9+ , L !+

+

#, , 40x

.

/ /91 / !"#$ /99

6 9 , &

ET/AO

v w ! !+ , #+

& " q 91 .

' #1 ' , 6 A

#, , .x40

6 9 q 91 , : &

, #E

& " F T D B .

B

hV : Ai9D 20 9+ , L !+

+ ,

#, 40 x

. 6 9 q 91 , 0+) $ $ &

, F # >

T D B .

C

9 hV : Ai9D 10 9+ , L !+

+ ,

#, 40x

. ' &

& " B =)

6 9 91 , : &

#E q AE4

6 9 , , &

"+, # > q 91 A

. 9 D

hV :

Ai9D 5 9+ , L !+

+

#, , 40x

.

&I 1 . / &J K $& @ L " K K?

! " %

& ' ( )

* ! " + , "

-. /

) 1 2 3 ' ,

(

4 % 0 200

18.5 200 %

0.5

24 % 200

1

51 % 200

2

52 200 %

5

0 20 40 60 80 100 120

5 2 1 0.5 0

Concentration µg/ml

Viability % BCL1

24 hr 48 hr 72 hr

M%

1.

/ " B &J $

- 5 BCL1 24 48، 72 OP 6/7 8/ 9

"

. MTT / & "

10⁴

× 2 L ' /K

96

% U

@0

$ V &D

6/7 8/ 9

) M% B F (

" & B , P 5 & MTT

24 Y 48 72 Z +% <

&D . [\I ]%

0 $ , %

100 B , 6%

$ , 0 " &J &D -?

&D ) M< F (

Y

p<0.05 B , 6%

$&D .

! "

IC50

, geX ? E) : q$

A

# x %

A4) : 9D 4+4 ) _ , MTT

i

:

TX )

50 _ Z 6 9 + &

E / )

8 .(

6 X 2 , ‡ , •

i9D = IC50

TX )

50

_ Z + ,

A4) K & MTT

E / ' : A

. ' &

& " B

1. half maximal inhibitory concentration

6 9 B &

T+ F "+,

AE4 e$

,

z , 6 9

=+EB &

' #+

IC50

, - , A ' : , 4 .

& " , 50

_

f ˆ 9 T+

24 Ad 6 9

6 ; &

: +, , A;

23 +

! 9+ L : +

J : +

;

; Ai9D 5

/ 9

!+

L

9+

+ F+ & ) f ˆ T+ ' #+

72 Ad

M

<

6 9 F "+, #+ +Y!) 6 ; &

T+

e$

72 ' " Ad

&

.

/ /91 / !"#$ /99

M%

2 . " B &J /

F %

- 5 24 48، 72

<

OP 6/7 8/ 9

"

. MTT / & "

10⁵

× 2 L ' /K

96

% U

@0

$ 6/7 V &D 8/ 9

) F

M% B (

" & B , P 5 & MTT

24 Y 48 72 Z +% <

&D .

0 $ , % [\I ]%

100

B , 6%

$ , 0 " &J &D -?

&D ) M< F (

Y p<0.05 B , 6%

$&D .

M%

3 .

" B &J /

F ! "

24 48، 72 OP <

6/7 8/ 9

"

. MTT

M%

4 . " &J ,

/ $ F ;

/ $ ! " F ، 48 BCL1

<

- @%

6/7 &

- / $ O " "

! " F ،

@ Y

D L 0 "

.

&I 2 . / $ IC50

،BCL1 / %

! "

/ Z , O F

Cancerous cell line (BCL1)

Proliferating normal cells

normal cells Resting

Treatment time

4.19 8.18

23.15 24 hr

3.76 8.48

10.59 48 hr

3.12 2.94

9.52 72 hr

/ /91 / !"#$ /99

6 9

=) , &

10⁵

× 2 0& Q &

A+9$

96

A"

Ai9D , l = &

N9 @ ) / .1 (

A4) 1 >

MTT

: =, 24 48 72 L M Ad .

geX F+% +

6 = 6 100

1 i

_

+., +4+4 ) 4 . ' , &

) /

d (

p<0.05

1 i

A .

_F + % ,

= ^ ‰

1Š :

! &

6 2012 V

) A1 , , F+ 4 ! : $ $ ' #+ TVT

, .

' " • ,

+

' : , ]d F+ 4 !

: : $ $ . ,

6 9 +Y!) + 9X #+ ) &

) 6 ( ) (<

, F+ 4 ! V #+ % - = ^

&

N9 @

?Y B

Hella A549 SKOV3

,

& ( )

T+ ) ,

&

% , i . : $ $ ' #+ F++=)

, 0+4 ) V % &

, 4,

' " F+ 4 ! ' : )

9 ( .

6 1999

Nefić Ibrulj

V 0+4 )

4 ! A+ C , F+

, ^+/ ' &

- _

in

vitro

= ^ .

A+ C ( ^+/ ' &

X 6 9 F

Ai9D l = &

N9 @ &

m F+ 4 !

in vitro

>

A, V

A+ C >

Ai9D l = ^+/ ' &

&

) 05 / 0 ) 1 / 0 9+ , L !+

+ ( 1 >

: + A+ =1 0 /) '

<

; + >

i9D l = A

)x , &

) Ai9D &

5 / 0 1 20

9+ , L !+

+ ( A+ C +Y!) , &

B

=

&

<

F+ W &

Ai9D †d , x , &

: U : ) Ji : 1 +$

?! ,

6 9 Ji e &

) 10

<

( ' )

1. Ozalp

= ^ E

+ F V ,

,

9 6 L M =+EB +Y!) 6 ; &

"

, . = ^

, X ) , b ; ,IC50

A 6 9 , &

6 /B ' )

M+

, F+ 4 ! V A1

6 9 6 9 6 &

6 /B A; 6 ; &

Ai9D ' : , 4, A

, . A e ' : =

q$

: 72 V F )x , Ad , 0+4 )

6 9 , 6 &

6 /B +Y!) 6 ; k ‚

' )

& "

F+ &

Ai9D B )x , &

20 10

L !+

9+ , 0+4 ) V +

AE4 "+,

, Ai9D F+ $ &

) X ) ,.

, ,IC50

A , 4, V #+ 9 ,

' :

Ai9D

' )

& "

F

Ai9D 0+4 ) &

9 , e V , "

AE4 6 9 , F+ $ 6 /B &

]Y ) 5 , L !+

9+

+ A 4 . .

6 9 , 9 IC50

&

6 ; +) : q$ +Y!) 72

' " d

&

6 9

&

Y!) 6 ; T+ +

9 : ) e$

< 4&

A+ C A ) ' : #1 , U>

&

9 Z ? / "+, +Y!) 6 ; .

:

&

+ L C b

F+ 4 ! C

4

<

J(

) F

!9 d

F

&

. : $ $ A

F _ ‚

?!+

9 A , F+ , )

&

?‚

†d ,

' + # +9$

6 , ) !+

&

<

F+ W &

F+ 4 ! :

' + # +9$

& ; : F+ , )

,

6 , ) !+

&

#+

+ 9X :

M

6 , ) !+

&

?+!") p

: + A

, i 9

F

? d , 0+

$

6 , ) !+

&

J+4.) 9

? @ Q

9 N>

. : $ $

A ) 6 (<

: 1 B A+ C

&

: , X J 4+

4&

+Y!)

#X 0C x [ $ A

) 4 (.

, X ) ,

J4+ !

? d F+4 !

Q 9

/ /91 / !"#$ /99

f &

+ 6 9

&

"+, f &

>

+

"+, J+4.) ,

&

"+, k ‚

V

0+4 ) F

:

&

, 6 9

&

6 =1 6 ;

+Y!) A+ C

&

6 ; +Y!) b

, i .

F - = ^ )

: 1 B A(X

F ' - C)

&

' + 6 9 6 ; +Y!)

=+EB

6 9 6 ; +Y!) B

+C , :

f B

% , l d

; F

&

,

6 9

&

6 =1 6 ;

[ $ J 4+

F : .

`

1. Kova E. Investigation of the proliferation, apoptosis/necrosis, and cell cycle phases in several human multiple myeloma cell lines. comparison of Viscum album QuFrF extract with Vincristine in an in vitro model. The Scientific World Journal . 2010 ; 10 : 311–320

2. Tait S.W.G, Green D.R . Mitochondria and cell death:

outer membrane permeabilization and beyond . Nature Reviews. 2010 ; 11 : 621 -632

3. Magalska A, Sliwinska M, Szczepanowska J, Salvioli S, Franceschi C, Sikora E . Resistance to apoptosis of HCW-2 cells can be overcome by curcumin- or vincristine-induced mitotic catastrophe . Int. J. Cancer . 2006 ;119: 1811–1818.

4. Abbas A.K, Lichman A, PilIa S . Cells and tissues of the adaptive immune system . In : Cellular and molecular immunology . Elsevier. 2007 ; 52

5. Krishna R ,Webb M, Onge G, Mayer L . Liposomal and nonliposomal drug pharmacokinetics after administration of liposome-encapsulated vincristine and their contribution to drug tissue distribution properties . The journal of pharmacology and experimental therapeutics . 2001; 298 :1206–1212

6. Gursel O, Sari E, Altun D, Atay AA, Akin R . Vincristine induced unilateral ptosis in a child. Pediatr Neurol. 2009;41:461-463

7. Yaraee R, Ghazanfari T, Eghtedardoost M, Rajabi M, Naseri M . The effect of MS14 on innate and cellular

immune responses in BALB/c mice.

Immunopharmacology and Immunotoxicology. 2011;

33(3): 509–514

8. Babykutty S, Padikkala J, Sathiadevan P, Vijayakurup V, Abdul AzisT,Srinivas P, Gopala S . Apoptosis induction of centella asiatica on human breast cancer cells .Babykutty et al., Afr. J. Trad. CAM. 2009; 6 (1): 9 - 16

9. Bian Y, Yang H, Yang Z, Gao F, Zhang N, Xiao C.

Amlodipine treatment prevents angiotensin II-induced human umbilical vein endothelial cell apoptosis. Archives of medical research . 2011;42:22-27

10. KosmiderB,Zynerb E, Osiecka R, Ochocki J . Induction of apoptosis and necrosis in A549 cells by the cis-Pt(II) complex of 3-aminoflavone in comparison with cis-DDP.

Mutation Research . 2004 ; 583 : 61–70

11. Lewis L. N. Chemical catalysis by colloids and clusters.

Chem. Rev. 1993 Dec; 93 (8): 2693–2730

12. Ozalp G, Zik B, Bastan A, Peker S, Ozdemir-Salcı E, Bastan I, Darbaz I, Salar S, Karakas K. Vincristine modulates the expression of Ki67 and apoptosis in naturally occurring canine transmissible venereal tumor (TVT). Biotechnic and histochemistry 2012;in press:1-6 12. Ying-jia Zhong, et al. Crocetin induces cytotoxicity and

enhances vincristine-induced cancer cell death via p53- dependent and -independent mechanisms. Acta Pharmacologica Sinica 2011; 32: 1529-1536

14. Nefić H, Ibrulj S . Genotoxic effect of the antitumor agent vincristine sulfate on human peripheral blood lymphocytes in vitro . Medicinski arhiv . 1999;53(4):185-8

Daneshvar(medicine) shahed University/ 19th year/2012/NO.99

Medicine

Scientific-Research Journal of Shahed

University Seventeenth Y ear,

No.99 June, Julay

2012

Comparing the efffect of vincristine on proliferating normal lymphocytes and BCL1 cell line

Mohaddese Shahhosseini¹, Sussan Kabudanian Ardestani², Roya Yaraee^3*

1- Department of Immunology, Faculty of Medicine, Shahed University, Tehran, Iran

2- Department of Biochemistry, Institute of Biochemistry and Biophysics, University of Tehran

3- Department of Immunology, Faculty of Medicine, Shahed University, Tehran, Iran

E-mail: ryaraee@yahoo.com

Asbtract

Background and Objective: Proliferation and apoptosis of lymphocytes are essential parts of the immune system. Most anti-cancer chemotherapy drugs such as vincristine target cell cycle and induce apoptosis in cancer cells .In other words, dividing cells undergo more apoptosis, which may also include the normal proliferating lymphocytes responsive to malignancies as well.

Materials and Methods: In this study, the effect of different concentration of vincristine at three different time periods on resting and proliferating spleen lymphocytes were evaluated and compared with the effect of the drug on mouse lymphoma cell line BCL1. The cytotoxicity of vincristine was determined by MTT assay and IC50 was calculated for all periods. The cells were also stained with double staining acridine orange and ethidium bromide and were observed with fluorescence microscope and the percentage of apoptotic cells were determined.

Results: MTT results showed that vincristine at concentrations of 10 and 20 g/ml caused cell death in both resting and proliferating lymphocytes, but concentrations<5 g/ml did not show any significant cytotoxic effect, while concentrations of 2 and 5 g/ml showed significant cytotoxic effect on BCL1 cells. The percentage of the apoptotic cells which were affected by different concentrations of the drug was proportional in the two methods, i.e. fluorescence microscope and MTT assay.

Conclusion: Vincristine has a strong cytotoxic effect in tumor cells and its toxic effect in normal cells is highly dependent on time and the activation of the cells.

Key words: Apoptosis, Proliferation, BCL1 (lymphoma cell line), Vincristine, Spleen

Received: 25/4/2012 Last revised: 3/7/2012

Accepted: 10/7/2012