PHA-767491联合TRAIL抑制肝癌细胞Bel-7404增殖  

Combination of PHA-767491 with TRAIL Inhibits Proliferation of Hepatocellular Carcinoma Bel-7404 Cells

作　　者：李伟[1] 李华美[1] 沈红强[1] 楼金吐[1] 彭朝阳[1] 董敖[1] 尚世强[1] 

机构地区：[1]浙江大学医学院附属儿童医院,实验检验中心,杭州310003

出　　处：《中国生物化学与分子生物学报》2013年第10期941-947,共7页Chinese Journal of Biochemistry and Molecular Biology

基　　金：浙江省医药卫生一般计划项目(No.2009B097);浙江省自然科学基金项目(No.Y2100891)~~

摘　　要：将小分子抗癌药物PHA-767491和携带TRAIL基因的非复制型腺病毒(Ad-TRAIL)共同作用于肝癌细胞Bel-7404,探讨PHA-767491联合TRAIL蛋白对肝癌细胞增殖的协同抑制作用及其作用机理.MTT法检测细胞存活率,Hoechst 33342荧光检测细胞凋亡现象和流式细胞仪检测细胞凋亡水平.结果表明,PHA-767491联合Ad-TRAIL抑制Bel-7404细胞增殖的能力显著优于单一用药.Western印迹进一步分析蛋白表达水平显示,PHA-767491可以通过下调抗凋亡蛋白Mcl-1和Xiap的表达,从而显著增强TRAIL蛋白诱导Bel-7404细胞凋亡的能力;PHA-767491联合AdTRAIL处理Bel-7404细胞后,不仅Bel-7404细胞凋亡水平显著增加,并且伴随着PARP和Caspase3的大量剪切.本研究证实,PHA-767491和TRAIL的联合使用对抑制肝癌细胞Bel-7404增殖表现出了显著的协同效应,为今后癌症药物的联合治疗提供了新的思路.To explore the coordinating repression to hepatocellular carcinoma cell proliferation by PHA- 767491 combining TRAIL and the inhibiting mechanism, small molecule anti-cancer drug PHA- 767491 and replication-deficient adenoviral vector carrying TRAIL （Ad-TRAIL） were used jointly to work on Bel- 7404 cell. By testting cell survival rate by MTT and observing apoptosis phenomenon via Hoechst 33342 fluorescence staining and apoptosis level by flow cytometry, 767491 with Ad-TRAIL promoted the growth inhibition of. the resuhs showed that combining PHA- Bel-7404 cell compared with either PHA- 767491 or Ad-TRAIL alone. Western blots analysis showed that PHA-767491 enhanced to induce apoptosis of TRAIL by down-regulating Mcl-1 and Xiap expression; PHA-767491 combined with Ad- TRAIL induced remarkable apoptosis and cleavage of caspase 3 and PARP in Bel-7404 cells. In conclusion, combination therapy of PHA-767491 and TRAIL exerted a synergistic anti-tumor effect on Bel-7404 cells, it will provide a new idea for combined treatment of cancer.

关 键 词：PHA-767491 TRAIL 协同效应 凋亡 BEL-7404 

分 类 号：Q78[生物学—分子生物学]