摘要
目的探讨苦参碱(matrine,Mat)诱导SKOV3细胞凋亡及可能的作用机制。方法四甲基偶氮唑蓝(MTT)法检测不同浓度Mat(0.4、0.8、1.2、1.6、2.0g·L-1)处理24、48、72h对SKOV3细胞增殖作用;流式细胞仪检测细胞的凋亡率,RT-PCR检测Survivin mRNA表达,Western blot检测Sur-vivin和Caspase-3蛋白的表达。结果一定浓度范围的Mat对SKOV3细胞的增殖均有抑制作用,且呈量效和时效关系,24、48、72h的IC50分别为3.38、1.57、1.13g·L-1;0.8、1.6g·L-1Mat作用48h后SKOV3细胞凋亡率分别为(11.44±0.56)%、(17.68±0.98)%,与空白对照组(4.85±0.31)%比较差异具有显著性(P<0.05)。RT-PCR及Western blot分析表明药物处理后SKOV3细胞的Survivin mRNA和蛋白表达均明显下调,而Caspase-3蛋白表达增加(P<0.05)。结论 Mat能抑制卵巢癌SKOV3细胞的增殖,促进其凋亡,其作用机制可能与下调Survivin表达和提高Caspase-3活性有关。
Aim To explore the effect of matrine（ Mat） on the apoptosis of ovarian cancer SKOV3 cells and its related mechanism. Methods SKOV3 cells were treated with different concentrations（ 0. 4,0. 8,1. 2,1. 6, 2. 0 g·L -1） of Mat for 24,48,72 h. The inhibitory rate of mat on SKOV3 cell proliferation was determined by MTT assay,and cell apoptosis was evaluated by flow cytometry （ FCM） ; the expression of survivin mRNA was measured by RT-PCR and the expression of survivin and Caspase-3 protein was measured by Western blot. Results Different concentrations of Mat had certain inhibitory effect on SKOV3 cell proliferation,in a dose-and tine-dependent manner. The half maximal inhibitory concentration （ IC50 ） of 24,48,72 h were 3. 38,1. 57,1. 13 g·L -1 respectively; the cell apoptotic rate of treated with 0. 8,1. 6 g·L -1 Mat for 48 h, was（ 11. 44 ± 0. 56） % ,（ 17. 68 ± 0. 98） % respective-ly,which had statistical significance compared with the control group（ 4. 85 ± 0. 31） % （ P〈0. 05） . The expressions of survivin in SKOV3 cells were down-regulated markedly at both mRNA and protein levels,but Caspase-3 protein was up-regulated（ P〈0. 05） . Conclusion Mat can inhibit the proliferation and induce the apoptosis of SKOV3 cells,and its mechanism might be related to down-regnlating the expression of survivin and increasing Caspase-3 activity.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2010年第8期1104-1107,共4页
Chinese Pharmacological Bulletin
基金
重庆市科委自然科学基金资助项目(No2005BB5314)
