摘要
目的探讨组蛋白去乙酰化酶抑制剂曲古菌素A与蛋白酶体抑制剂硼替佐米单独及联合应用对人卵巢癌细胞株SKOV3存活率和凋亡率的影响。初步探讨两种药物联合应用对诱导SKOV3细胞凋亡具有的协同作用。方法曲古菌素A、硼替佐米单独或者联合应用于卵巢癌细胞后,用四甲基偶氮唑蓝(MTT)比色法测定细胞增殖活性,并计算细胞存活率,Annexin-V/PI法流式细胞仪检测细胞凋亡率,蛋白质免疫印迹法(Western blot)检测相关蛋白表达水平。通过检测Caspases-3的活性及其底物多聚ADP核糖聚合酶(PARP)的表达水平进一步说明不同用药组诱导细胞凋亡的情况。结果联合用药组诱导的细胞凋亡率和单独用药组比较,差异有统计学意义(P<0.05)。几种凋亡相关蛋白Bcl-2、Mcl-1和Bcl-XL,在联合用药组的表达显著低于单独用药组。在相同的时间点,联合用药组Caspase-3的活性和单独用药组比较,差异有统计学意义(P<0.001)。结论低剂量的曲古菌素A和硼替佐米联合作用人卵巢癌细胞系能诱导凋亡,并且这种作用远远强于相同剂量单独用药引起的凋亡。这两种药物的联合应用可能成为人卵巢癌化疗中的新方案。
Objective To investigate the apoptotic effect exerted on human ovarian cancer cells by both the histone deacetylase inhibitor Trichostatin A（TSA）and the proteasome inhibitor PS-341 and their synergistic effect.Methods Methyl thiazolyl tetrazolium（MTT）assay was applied to examine the cell viability,Annexin-V/PI apoptosis detection kit was used to determine the apoptosis rate of different groups.Besides,Western blot was introduced to evaluate the expression levels of Bcl-2、Mcl-1 and Bcl-XL.To furtherly evaluate apoptosis in different groups,the activity of Caspase-3 and expression level of PARP was detected.Results The cell apoptosis rates which were caused by TSA/PS-341 regimen or TSA,PS-341 alone were different（P0.05）.Western blot showed that after the treatment of TSA and PS-341 combination,Bcl-2、Mcl-1 and Bcl-XL were down-regulated.Exposure to TSA/PS-341 induced the activity of Caspase-3 and the expression of PARP after 48 h of treatment,and there were significant differences compared with the treatment of drug alone（P0.001）.Conclusion Low-dose TSA and PS-341 synergistically induce cytotoxicity in ovarian cancer cells,and the effect was much more significant than single drug treatment groups.TSA/PS-341 regimen may represent a potential novel therapeutic strategy for ovarian cancer.
出处
《肿瘤防治研究》
CAS
CSCD
北大核心
2010年第8期889-893,共5页
Cancer Research on Prevention and Treatment
基金
国家973重点基础科学基金资助项目(2002CB513107)
关键词
曲古菌素A
硼替佐米
凋亡
协同作用
卵巢癌细胞
Trichostatin A
PS-341
Apoptosis
Synergistic interaction
Ovarian cancer cells
