半胱氨酸酶-3在依托利酸诱导肝癌细胞凋亡中的作用  

Significance of Caspase-3 in etodolac-induced apoptosis of SMMC7721 cell line

作　　者：周喜乐[1] 郑树森[1] 谢海洋[1] 

机构地区：[1]浙江大学医学院附属第一医院肝胆胰外科,杭州310003

出　　处：《中华实验外科杂志》2004年第5期541-543,共3页Chinese Journal of Experimental Surgery

基　　金：浙江省科技厅资助项目 (0 2 1 1 0 72 4 1 )

摘　　要：目的　探讨选择性环氧合酶 (COX)抑制剂依托利酸 (etodolac)诱导肝癌SMMC772 1细胞凋亡的分子机制。方法　采用流式细胞术、DNA琼脂糖凝胶电泳法测定细胞凋亡情况 ;West ernblotting法检测不同浓度etodolac处理后凋亡相关蛋白Bcl 2、Bax表达的变化 ;并以流式细胞术检测半胱氨酸酶 3 (Caspase 3 )酶活性的变化。 结果　流式细胞术显示etodolac(0、0 .2 5、0 .5 0、1.0 0、2 .0 0mmol/L)作用SMMC772 1细胞 48h后 ,对照处理组 (0mmol/L)没有出现凋亡峰 ,其余各组(0 .2 5、0 .5 0、1.0 0、2 .0 0mmol/L)出现明显的凋亡峰 ,其凋亡率分别为 (16.3± 3 .1) %、(19.9±3 .6) %、(2 2 .9± 3 .2 ) %、(3 1.2± 3 .3 ) % (P <0 .0 0 1) ;DNA琼脂糖凝胶电泳出现明显的DNALad der ;不同浓度etodolac处理后凋亡相关蛋白Bcl 2表达下降 ,Bax表达增加 ;随着etodolac处理浓度的增加 ,表达活性Caspase 3的细胞百分率分别为 (3 .61± 0 .3 2 ) %、(2 .93± 0 .15 ) %、(10 .2 9±0 .3 9) %、(2 7.3 3± 1.2 8) %、(5 7.40± 1.69) % (P <0 .0 0 1)。结论　选择性COX 2抑制剂可能通过调节Bcl 2、Bax蛋白表达活化Caspase 3 ,从而诱导肝癌SMMC772 1细胞凋亡。Objective To investigate the possible role of Bcl-2,Bax and Caspase-3 in etodolac-induced apoptosis of liver tumor SMMC7721 cell line.Methods Cell apoptosis was determined by flowcytometry analysis using PI staining,DNA laddering.The expression of Bcl-2 and Bax protein was detected by Western blotting.Caspase-3 activity was evaluated by active Caspase-3 apoptosis kit with flow cytometry.Results Selective COX-2 inhibitor etodolac stimulated apoptosis in liver tumor SMMC7721 cell line significantly.Flowcytometry revealed the apoptotic rate was (16.3±3.1)%,(19.9±3.6)%,(22.9±3.2)%,(31.2±3.3)% with different concentrations of etodolac (0.25,0.50,1.00,2.00 mmol/L),while the apoptotic peak did not appear in the control group (0 mmol/L,P <0.001).The expression of Bax protein was up-regulated while Bcl-2 protein was down-regulated,and the percentage of the cells with active Caspase-3 was (3.61±0.32)%,(2.93±0.15)%,(10.29±0.39)%,(27.33±1.28)%, (57.40± 1.69)% separately with the increases of etodolac concentrations [(0,0.25,0.50,1.00,2.00) mmol/L, ( P < 0.001)]. Conclusion Selective COX-2 inhibitor etodolac may activate Caspase-3 by regulating the expression of Bcl-2,Bax protein,which causes apoptosis of SMMC7721 cells.

关 键 词：半胱氨酸酶-3 依托利酸 诱导 肝癌 细胞凋亡 CASPASE-3 

分 类 号：R735.7[医药卫生—肿瘤]