天花粉蛋白联合siPD-1促进卵巢癌细胞凋亡及机制研究  

Research on trichosanthin combined with siPD-1 promotes apoptosis of human ovarian cancer cells and its mechanism

作　　者：邹立君[1] 胡律江(指导)[2] ZOU Li-Jun;HU Lyu-Jiang(Jiangxi College of Traditional Chinese Medicine,Fuzhou 344000,China)

机构地区：[1]江西中医药高等专科学校,抚州344000 [2]江西中医药大学附属医院,南昌330000

出　　处：《中国免疫学杂志》2021年第24期2981-2985,共5页Chinese Journal of Immunology

基　　金：江西省教育厅科学技术研究项目(170707);江西省卫生健康委中医药科研课题项目(2017A322)。

摘　　要：目的:分析天花粉蛋白(TCS)、TCS和siPD-1联合用药对卵巢癌细胞凋亡的影响,并探讨其作用机制。方法:将卵巢癌SKOV3细胞接种于6孔板(1×10^(5)个/孔),并分为3组,空白对照组:12 h后更换培养液,无任何药物处理,TCS处理组:12 h后加入含不同浓度TCS(20、40、60、80、100μg/ml)的培养基,TCS+siPD-1联合处理组:12 h后加入含不同浓度TCS(20、40、60、80、100μg/ml)和不同终浓度siPD-1(60、80、100、120、150 nmol/L)的培养基,培养至24 h、48 h时收集细胞,流式细胞术检测细胞凋亡,RT-PCR及ELISA检测凋亡相关信号表达及酶活性。结果:与空白对照组相比,TCS处理后的SKOV3细胞凋亡率显著提高,且呈时间和剂量依赖性(P<0.05);与TCS处理组相比,TCS+siPD-1处理组细胞凋亡率显著上升(P<0.05)。不同浓度TCS处理组、TCS+siPD-1处理组细胞Caspase-3、Caspase-6、Caspase-8、Caspase-9 mRNA表达上升,且呈时间和剂量依赖性(P<0.05),TCS+siPD-1处理组Caspase-3、Caspase-6、Caspase-8、Caspase-9 mRNA表达显著高于TCS处理组(P<0.05)。不同浓度TCS处理组、TCS+siPD-1处理组48 h后,细胞Caspase-3酶活性显著上升,且呈剂量依赖性。结论:TCS和siPD-1联用可促进卵巢癌细胞凋亡,其机制可能为通过调节Caspase-3相关凋亡通路级联反应实现的。Objective:To analyze effects of Trichosanthin(TCS),TCS combined with siPD-1 on apoptosis of ovarian cancer,and to investigate its mechanism.Methods:Ovarian cancer cells SKOV3 were inoculated into six-well plate(1×10^(5) cells/well),and divided into 3 groups,blank control group:culture medium was changed after 12 h without any drug treatment.TCS group:medium containing different dosage of TCS(20,40,60,80,100,120μg/ml)was added after 12 h.TCS+siPD-1 group:medium containing different dosage of TCS(20,40,60,80,100,120μg/ml)and different dosage of siPD-1(60,80,100,120,150 nmol/L)was added 12 h later.Cells were collected at 24 h and 48 h after culture.Cell apoptosis was detected by flow cytometry,and expressions of apoptosis related signal and enzyme activity were detected by RT-PCR and ELISA.Results:Compared with blank control,apoptosis rate of SKOV3 after TCS treatment was significantly increased,which was in a time and dose dependent manner(P<0.05).Compared with TCS group,cell apoptosis rate of TCS+siPD-1 group was increased significantly(P<0.05).mRNA expressions of Caspase-3,Caspase-6,Caspase-8 and Caspase-9 in cells treated with TCS and siPD-1 at different concentrations were increased in a time-dependent and dose-dependent manner.mRNA levels of Caspase-3,Caspase-6,Caspase-8 and Caspase-9 in TCS+siPD-1 group were significantly higher than those in TCS group(P<0.05).After 48 h,Caspase-3 activity was increased significantly in TCS treatment group and TCS+siPD-1 treatment group in a dose-dependent manner.Conclusion:TCS combined with siPD-1 synergistically can induce apoptosis of ovarian cancer cells,whose mechanism may by regulating Caspase-3 mediated cascade reaction.

关 键 词：天花粉蛋白 siPD-1 卵巢癌 细胞凋亡 CASPASE 

分 类 号：R737.31[医药卫生—肿瘤]