20(s)-原人参二醇对体外培养Siha细胞caspase-3表达激活的影响  

Effects of 20(s)-proto-panaxdiol on caspase-3 expression and activation in Siha cells in vitro

作　　者：赵丽晶[1] 许多[1] 程宏[1] 梁作文[1] 鲁育铭[1] 韩向北[1] 郭亚雄[1] 赵丽娟[1] 董妍[2] 

机构地区：[1]吉林大学白求恩医学院病理生理学教研室,吉林长春130021 [2]美国杜兰大学细胞生物学教研室,美国新奥尔良la70062

出　　处：《吉林大学学报（医学版）》2011年第1期26-29,I0001,共5页Journal of Jilin University：Medicine Edition

基　　金：吉林省人民政府人才开发基金资助课题(2006JD02)

摘　　要：目的:观察20(s)-原人参二醇(PPD)对体外培养人宫颈癌Siha细胞中caspase-9和caspase-3转录表达的影响,以及caspase-3的活化形式cleaved caspase-3含量的变化,阐明其诱导Siha细胞凋亡的机制。方法:体外培养人宫颈癌Siha细胞,将其分为阴性对照组(乙醇)和实验组(20μg.L-1 PPD),分别用乙醇和PPD处理体外培养的宫颈癌Siha细胞,48h后流式细胞仪检测细胞的凋亡峰,半定量RT-PCR、Western blotting和免疫细胞化学染色方法检测PPD处理后Siha细胞中caspase-9和caspase-3基因的转录与表达水平。结果:与阴性对照组比较,20μg.L-1 PPD处理48h后,Siha细胞凋亡率增加(P<0.05),Siha细胞中caspase-9与caspase-3转录水平升高(P<0.01),表达上调(P<0.01),caspase-3的活化形式cleaved caspase-3含量增加(P<0.01)。细胞免疫化学检测可见实验组细胞出现棕色颗粒。结论:PPD可促进人宫颈癌Siha细胞凋亡,其机制与激活caspase家族级联反应有关。Objective To study the effects of 20（s）-proto-panaxdiol（PPD）on transcription and protein expressions of caspase-9 and caspase-3,and content of cleaved caspase-3 in Siha cells in vitro,and clarify the mechanism of its apootosis action.Methods Siha cells cultivated in vitro were treated with alcohol（negative control group）and 20 μg·L-1 PPD（20 μg·L-1 PPD group）,respectively.The apoptosis was detected by FCM 48 h after treatment.The transcription and protein expressions of caspase-9 and caspase-3 in siha cells were analyzed by RT-PCR,Western blotting and immunocytochemical staining.Results Compared with negative control group,after treated with 20 μg·L-1 PPD for 48 h,the apoptotic rate of Siha cells was increased（P0.05）,the transcription and protein expressions of caspase-9 and caspase-3 were up-regulated（P0.01）,and the content of cleaved caspase-3 was increased（P0.01）.Cell immunochemistry detection showed the brown particles in the experimental cells in 20 μg·L-1 PPD group.Conclusion PPD may induce the apoptosis of Siha cells,and the mechanism may be related to the up-regulation and activition of caspase family.

关 键 词：20(S)-原人参二醇 SIHA细胞 细胞凋亡 CASPASE-3 CASPASE-9 基因表达 

分 类 号：Q75[生物学—分子生物学] R737.3[医药卫生—肿瘤]