Anti2leukemia effect of resveratrol of Polygonum cuspidatum exerts and possible molecular mechanism

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1 ( ) J ournal of Xiπan J iaot ong U niversity (Medical Sciences) V ol. 29 No. 3 J un , 2, 3, 4, 2 (1., ;2., ;3., ; 4., ) : M TT ; H E ; Western Blot2 ting Bcl22 ; p2sta T3 2 3 STA T3, STA T3, : ; ; :R967 :A : (2008) Anti2leukemia effect of resveratrol of Polygonum cuspidatum exerts and possible molecular mechanism Li Tan 1, Fan Guixiang 2, Wang Wei 3, Li Tong 4, Yuan Yukang 2 (1. Depart ment of Immunology, Medical College of t he Chinese Peopleπs Armed Police Force, Tianjin ; 2. Depart ment of Immunology and Pat hogenic Biology, Medical School of Xiπan Jiaotong U niversity, Xiπan ; 3. Department of Nep hrology, t he Affiliated Hospital of Medical College of t he Chinese Peopleπs Armed Police Force, Tianjin ; 4. Department of Ultrasonography, Xinjiang Armed Police General Hospital, Urumqi , China) ABSTRACT : Objective To explore t he anti2leukemia eff ects of resverat rol on t hree leukemia cell lines Hu T278, J urkat and L 1210 in vivo. Methods M TT colorimet ry was used t o measure t he tumor cell growt h rep ression rate. The level of Bcl22 was measured by Wester n blotting. The cell cycle of cells was analyzed by flow cyt omet ry. The exp ression of p2s TA T3 was determined by immunohist ochemistry. Results Our findings indicated t hat resverat rol could inhibit p rolif eration, induce ap op t osis, and influence cell cycle of Hu T278, J urkat and L 1210 cells in a dose2 and time2dependent manner and reduce t he exp ression of p2s TA T3 in vitro. Conclusion Resverat rol might have a chemot herapeutic p ote ntial f or leukemia of various sources and reduce t he exp ression of p2s TA T3 w hich p rovides experiment evidence f or its application in clinical t reat ment of leukemia. KEY WORDS : resverat rol ; ap op t osis ; leukemia,,,,,, [1],,, : : : (19782), ( ),,,. E2mail : com, (trans2resveratrol), [2],,, 3,,

2 3,,, ( %) ; M T T Sigma ; RPMI 1640 ( FCS) Gibco ; ( ) Bcl22 Labvision ; ( ) p2 STA T3 Santa Cruz ; Annexin V2FITC ; (L1210) ; T ( Hu T278) T (J ur2 kat, Clone E621) % ( ) CO2, 10 %( ) 1640,800 r/ min 5 min, , 1640 > 95 %, Nikon TMS2F ( HE) mol/ L h,,he 1. 4 M T T RPMI / ml, [3 ], 3 T h, 0. 5 %( ) RPMI h [4 ], / ml 3 ml,,5 %( ) CO2,37, 6 h PI FACSCalibur / ml,, 5 % ( ) CO2, 37, h FITC Annexin2 V PI FACS2 can 1. 7 Western Bcl22, 2 mg/ L SDS2PAGE, h Bcl22 ( TBST ),HRP2 ( TBST ),ECL, 2actin 1. 8 SP HuT278 J urkat L1210 p2stat 3 p2sta T ,DAB, mol/ L PBS, : ( - ) < 10 % ; ( + ) % ; ( 6 ) % ; ( 7 ) > 70 % 1. 9 SPSS13. 0 (one way ANOVA), ; Bonferroni t P < Hu T278 J urkat L mol/ L Hu T278 J urkat L h,,, ( 1) 2. 2 H E,,,,,,,,,,,,, ( 2) 2. 3,3 Hu T278 J ur2 kat L1210, - ( 3A - 3C), L1210, % ; Hu T278 J urkat % % 3, ( 3D), 3 T3 ( P < 0. 01), L1210 Hu T278,L1210 J urkat 12 h ( P < 0. 05), ; J urkat Hu T278, 48 h ( P < 0. 01)

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4 3,,, , G0 / G % %, S % % ( Hu T278) ;J urkat ;L1210 S : % %, S ( 4) 4 PI ( 6 h) Fig. 4 Resveratro2induced cell cycle perturbations and apoptosis in HuT278, J urkat and L1210 cells cultured for 6 h. Cells stained with PI A, C, E: control cells incubated without resveratrol ; B, D, F : HuT 78, Jurkat and L1210 cells treated with 50 mol/ L resveratrol, respectively mol/ L 12 h, 25 %L1210, 48 h, ( 5) ( 6), 100 mol/ L 12 h, HuT278 ( %) ;J urkat L % %,, HuT278 L1210 J urkat 2. 6 Bcl22 3 Bcl22, Hu T278 J urkat L1210 Bcl22, ( 7) 5 50 mol/ L L1210 Fig. 5 The morphology of apoptotic cells treated with 50 mol/ L resveratrol ( 20) A : 12 h ; B : 48 h mol/ L 12 h Fig. 6 The apoptotsis of different cells treated with 100 mol/ L resveratrol for 12 h by flow cytometric analysis A : Hu T278 cells ; B : J urkat cells ; C : L1210 cells

5 344 ( ) Hu T278 J urkat L1210 p2stat 3 p2stat 3, > 70 %,p2stat 3 ( P < 0. 05, 8) 7 48 h, Hu T278 J urkat L1210 Bcl22 Fig. 7 The relative expression level of Bcl22 in Hu T278, J urkat and L1210 cells treated with different doses of res2 veratrol for 48 h 3, L1210 Hu T278 J urkat, 3, L1210 Hu T278 J urkat, [5 ],,, [6 ], 3 Western Blot, Hu T278 J urkat L1210 Bcl22, 8 Hu T278 J urkat L1210 p2sta T3 Fig. 8 The expression of p2sta T3 in Hu T278, J urkat and L1210 cells A : p2sta T3 in Hu T278 ; B : p2sta T3 in Hu T278 treated wit h 100 mol/ L resveratrol for 24 h ; C : p2sta T3 in J urkat ; D : p2sta T3 in J urkat treated wit h 100 mol/ L resveratrol for 24 h ; E : p2sta T3 in L1210 ; F : p2sta T3 in L1210 treated wit h 100 mol/ L resveratrol for 24 h Bcl22, Bcl22,Bcl22 Bcl22, [728 ], Bcl22,, J A K/ STA T ( signal transducers and activators of transcription, ) [9210 ] STA T DNA,

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