extracellular caspase-8 dependent apoptosis on hela cancer cells and mrc-5 normal cells by icd-85 (venom derived peptides)

Authors

abbas zare mirakabadi dept. of venomous animals and antivenom production, razi vaccine and serum research institute, karaj, iran

ali sarzaeem dept. of venomous animals and antivenom production, razi vaccine and serum research institute, karaj, iran

abstract

background: our previous studies revealed an inhibitory effect of icd-85 (venom derived peptides) on mda-mb231 and hl-60 cell lines, through induction of apoptosis. the purpose of this study was to investigate apoptosis-induced mechanism on hela and mrc-5 cells by icd-85 through activation of caspase-8. methods: cell viability, cytosolic enzyme lactate dehydrogenase (ldh) and cell morphology were assessed under unexposed and icd-85 exposed conditions.caspase-8 activity was assayed by caspase-8 colorimetric assay kit. results: the results show that inhibitory concentration 50% (ic 50 ) value of icd-85 for hela cells at 24 h was estimated and found to be 25.32±2.15µg/ml. furthermore, treatment of hela cells with icd-85 at concentrations of 1.6×10 and 2.6×10µg/ml did not significantly increase ldh release. morphological changes in hela cells on treatment with icd-85 compared with untreated hela cells consistent with an apoptotic mechanism of cell death, such as cell shrinkage which finally results in the generation of apoptotic bodies. however, when mrc-5 cells were exposed to icd-85, no significant changes in cell morphology and ldh were observed at concentrations below 2.6×10µg/ml. also, the apoptosis-induction mechanism by icd-85 on hela cells was found through activation of caspase-8 and the activity of caspase-8 in hela cells was 1.5 folds more than its activity on mrc-5 cells. conclusion: therefore, the apoptosis-induced mechanisms by icd-85 are through activation of caspase-8 and concerning the least cytotoxic effect on mrc-5 cells, icd-85 may be used as anticancer compound to inhibit growth of cancer cells.

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Journal title:
iranian journal of cancer prevention

جلد ۵، شماره ۴، صفحات ۱۹۴-۲۰۲

Keywords
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o m e r a s e a c t i v i t y i n h u m a n c a n c e r s . e u r j c a n c e r . 1 9 9 7 ' , 3 3 , 7 8 7 , ' 9 1 . 3 9 . e b i s u n o s ' , ' i n a g a k i t ' , ' k o h j i m o t o y ' , ' o h k a w a t . t h e c y t o t o x i c e f f e c t o f f e r o x a c i n a n d c i p r o f o x a c i n o n t r a n s i t i o n a l c e l l c a r c i n o m a i n v i t r o . c a n c e r . 1 9 9 7 ' , 8 0 , 2 2 6 3 , ' 7 . 4 0 . g ü r b a y a ' , ' g a r r e l c ' , ' o s m a n m ' , ' r i c h a r d m j ' , ' f a v i e r a ' , ' h i n c a l f . c y t o t o x i c i t y i n c i p r o f o x a c i n ' , ' t r e a t e d h u m a n f i b r o b l a s t c e l l s a n d p r o t e c t i o n b y v i t a m i n e . h u m e x p t o x i c o l . 2 0 0 2 ' , 2 1 , 6 3 5 , ' 4 1 . 4 1 . w i j s m a n j a ' , ' d e k a b a n g a ' , ' r i e d e r m j . d i f f e r e n t i a l t o x i c i t y o f r e a c t i v e m e t a b o l i t e s o f c l y n d a m i c i n a n d s u l f o n a m i d e s i n h i v ' , ' i n f e c t e 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u s e s 5 0 p e r c e n t i n h i b i t i o n ( i 5 0 ) o f a n e n z y m a t i c r e a c t i o n . b i o c h e m p h a r m a c o l . 1 9 7 3 ' , 2 2 , 3 0 9 9 , ' 1 0 8 . 4 5 . n e u b i g r r ' , ' s p e d d i n g m ' , ' k e n a k i n t ' , ' c h r i s t o p o u l o s a a . i n t e r n a t i o n a l : u n i o n : o f p h a r m a c o l o g y c o m m i t t e e o n r e c e p t o r n o m e n c l a t u r e a n d d r u g c l a s s i f i c a t i o n . x x x v i i i . u p d a t e o n t e r m s a n d s y m b o l s i n q u a n t i t a t i v e p h a r m a c o l o g y . p h a r m a c o l r e v . 2 0 0 3 ' , 5 5 , 5 9 7 , ' 6 0 6 . 4 6 . f o t a k i s g ' , ' t i m b r e l l j a . i n v i t r o c y t o t o x i c i t y a s s a y s ' , ' c o m p a r i s o n o f l d h ' , ' n e u t r a l r e d ' , ' m t t a n d p r o t e i n a s s a y i n h e p a t o m a c e l l l i n e s f o l l o w i n g e x p o s u r e t o c a d m i u m c h l o r i d e . t o x i c o l o g y l e t t e r s . 2 0 0 6 ' , 1 6 0 , 1 7 1 , ' 7 . 4 7 . b a l d u z z i m ' , ' d i o c i a i u t i m ' , ' d e b e r a r d i s b ' , ' p a r a d i s i s ' , ' p a o l e t t i l . i n v i t r o e f f e c t s o n m a c r o p h a g e s i n d u c e d b y n o n c y t o t o x i c d o s e s o f s i l i c a p a r t i c l e s p o s s i b l y r e l e v a n t t o a m b i e n t e x p o s u r e . e n v i r o n m e n t a l r e s e a r c h . 2 0 0 4 ' , 9 6 , 6 2 , ' 7 1 . 4 8 . s a y e s c m ' , ' g o b i n a m ' , ' a u s m a n k d ' , ' m e n d e z j ' , ' w e s t j l ' , ' c o l v i n v l . n a n o ' , ' c 6 0 c y t o t o x i c i t y i s d u e t o l i p i d p e r o x i d a t i o n . b i o m a t e r i a l s . 2 0 0 5 ' , 2 6 , 7 5 8 7 , ' 9 5 . 4 9 . h u s s a i n s m ' , ' h e s s k l ' , ' g e a r h a r t j m ' , ' g e i s s k t ' , ' s c h l a g e r j j . i n v i t r o t o x i c i t y o f n a n o p a r t i c l e s i n b r l 3 a r a t l i v e r c e l l s . t o x i c o l o g y i n v i t r o . 2 0 0 5 ' , 1 9 , 9 7 5 , ' 8 3 . 5 0 . 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. 5 7 . z h a n g l ' , ' z h u h ' , ' t e r a i s h i f ' , ' d a v i s j j ' , ' g u o w ' , ' f a n z z ' , ' e t a l . a c c e l e r a t e d d e g r a d a t i o n o f c a s p a s e ' , ' 8 p r o t e i n c o r r e l a t e s w i t h t r a i l r e s i s t a n c e i n a d l d 1 h u m a n c o l o n c e l l l i n e . n e o p l a s i a . 2 0 0 5 ' , 7 , 5 9 4 , ' 6 0 2 5 8 . g e r a l d s s ' , ' x i n y i n g j i ' , ' w a n g w ' , ' l e i g h a c a l l a h a n . t h e e x t r i n s i c c a s p a s e p a t h w a y m o d u l a t e s e n d o t o x i n ' , ' i n d u c e d d i a p h r a g m c o n t r a c t i l e d y s f u n c t i o n . j a p p l p h y s i o l . 2 0 0 7 ' , 1 0 2 , 1 6 4 9 , ' 5 7 . 5 9 . l a g u i n g e l m ' , ' s a m a r a r n ' , ' w a n g w ' , ' e l ' , ' d e i r y w s ' , ' c o r n e r g . d r 5 r e c e p t o r m e d i a t e s a n o i k i s i n h u m a n c o l o r e c t a l c a r c i n o m a c e l l l i n e s . c a n c e r r e s e a r c h . 2 0 0 8 ' , 6 8 , 9 0 9 , ' 1 7 . 6 0 . a l f o n s l . a p o p t o s i s a n i n t r o d u c t i o n . r e v i e w a r t i c l e s . b i o e s s a y s . 2 0 0 3 ' , 2 5 , 8 8 8 , ' 9 6 . 6 1 . a r a k i s ' , ' m a s u d a s ' , ' m a e d a h ' , ' y i n g m j ' , ' h a y a s h i h . i n v o l v e m e n t o f s p e c i f i c i n t e g r i n s i n a p o p t o s i s i n d u c e d b y v a s c u l a r a p o p t o s i s i n d u c i n g p r o t e i n 1 . t o x i c o n . 2 0 0 2 ' , 4 0 , 5 3 5 , ' 4 2 . 6 2 . w a i d a j s ' , ' d o w d y s f . m o d u l a t i o n o f c e l l u l a r f u n c t i o n b y t a t m e d i a t e d t r a n s d u c t i o n o f f u l l l e n g t h p r o t e i n s . c u r r p r o t e i n p e p t s c i . 2 0 0 3 ' , 4 , 9 7 , ' 1 0 4 . 6 3 . b a l a s u b a s h i n i m s r i ' , ' k a r t h i g a y a n s ' , ' s o m a s u n d a r a m s t ' , ' b a l a s u b r a m a n i a n t ' , ' r u k k u m a n i r ' , ' v e n u g o p a l p m . f v p e p t i d e i n d u c e s a p o p t o s i s i n h e p 2 a 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