摘要
目的:探讨他克莫司(FK506)调节Yes相关蛋白(YAP)/转录共激活因子PDZ结合基序(TAZ)信号通路对卵巢癌细胞增殖、凋亡和侵袭的影响。方法:将对数生长期SKOV3细胞分为对照组、阴性对照(si-NC)组(转染si-NC质粒)、沉默YAP载体(si-YAP)组(转染si-YAP)、过表达质粒阴性对照(pcDNA-NC)组(转染pcDNA-NC)、过表达YAP质粒(pcDNA-YAP)组(转染pcDNA-YAP)、FK506组(500μg/L)、FK506+si-NC组(500μg/L FK506+转染si-NC质粒)、FK506+si-YAP组(500μg/L FK506+转染si-YAP)、FK506+pcDNA-NC组(500μg/L FK506+转染pcDNA-NC)、FK506+pcDNA-YAP组(500μg/L FK506+转染pcDNA-YAP)。CCK-8法检测细胞活力,流式细胞仪检测细胞凋亡,Transwell实验检测细胞侵袭,qRT-PCR检测YAP、TAZ mRNA表达,Western blot检测YAP、TAZ、基质金属蛋白酶2(MMP-2)、Bcl2-相关X蛋白(Bax)及细胞周期蛋白D1(CyclinD1)表达水平。结果:与IOSE80相比,SKOV3、OVCAR3、UWB1.289细胞中YAP、TAZ mRNA和蛋白表达水平显著增加(P<0.05)。与si-NC组相比,si-YAP组YAP、TAZ mRNA和蛋白表达水平、细胞活力、侵袭细胞数显著下降(P<0.05),细胞凋亡率显著增加(P<0.05);与pcDNA-NC组相比,pcDNA-YAP组YAP、TAZ mRNA和蛋白表达水平、细胞活力、侵袭细胞数显著增加(P<0.05),细胞凋亡率显著降低(P<0.05)。与对照组相比,FK506组细胞活力、侵袭数,YAP、TAZ mRNA表达,以及YAP、AZ、CyclinD1、MMP-2蛋白表达均显著降低,凋亡率及Bax表达显著增加(P<0.05);与FK506+si-NC组相比,FK506+si-YAP组细胞活力、侵袭数,YAP、TAZ mRNA表达,以及YAP、TAZ、CyclinD1、MMP-2表达显著降低,凋亡率及Bax表达显著增加(P<0.05);与FK506+pcDNA-NC组相比,FK506+pcDNA-YAP组细胞活力、侵袭数,YAP、TAZ mRNA表达,以及YAP、TAZ、CyclinD1、MMP-2表达显著增加,凋亡率及Bax表达显著降低(P<0.05)。结论:FK506通过抑制YAP/TAZ信号通路阻止SKOV3细胞增殖、侵袭,诱导其凋亡。
Objective:To investigate the impacts of tacrolimus(FK506)on the proliferation,apoptosis and invasion of ovarian cancer(OC)cells by regulating the Yes-associated protein(YAP)/transcriptional co-activator with PDZ-binding motif(TAZ)signal pathway.Methods:SKOV3 cells in logarithmic growth phase were divided into control group,negative control group(si-NC)(transfected with si-NC plasmid),silenced YAP vector group(transfected with si-YAP),overexpressed negative control(pcDNA-NC)group(transfected with pcDNA-NC),and overexpressed YAP plasmid(pcDNA-YAP)group(transfected with pcDNA-YAP),FK506 group(500μg/L),FK506+si-NC group(500μg/L FK506+transfected with si-NC plasmid),FK506+si-YAP group(500μg/L FK506+transfected with si-YAP),FK506+pcDNA-NC group(500μg/L FK506+transfected with pcDNA-NC),and FK506+pcDNA-YAP group(500μg/L FK506+transfected with pcDNA-YAP).CCK-8 method was applied to detect cell viability,flow cytometry was applied to detect cell apoptosis,Transwell experiment was applied to detect cell invasion,qRT-PCR was applied to detect YAP and TAZ mRNA expression,Western blot was applied to detect the expression levels of YAP,TAZ,matrix metalloproteinase-2(MMP-2),Bcl2 associated X protein(Bax),and CyclinD1.Results:Compared with IOSE80,the mRNA and protein expression levels of YAP and TAZ in SKOV3,OVCAR3 and UWB1.289 cells were obviously increased(P<0.05).Compared with si-NC group,the expression levels of YAP and TAZ mRNA and protein,cell viability and number of invasive cells in si-YAP group were significantly decreased(P<0.05),and the apoptosis rate was significantly increased(P<0.05).Compared with pcDNA-NC group,the expression levels of YAP and TAZ mRNA and protein,cell viability and number of invasive cells in pcDNA-YAP group were significantly increased(P<0.05),and the apoptosis rate was significantly decreased(P<0.05).Compared with the control group,the cell viability,invasion number,and the expressions of YAP,TAZ mRNA,YAP,TAZ,CyclinD1,MMP-2 in FK506 group were obviously reduced,the apoptosis rate and the ex
作者
李冬梅
孟维合
康聪
Li Dongmei;Meng Weihe;Kang Cong(Department of Pharmacy,Hengshui People's Hospital,Hengshui 053000;Department of Gynecology,Hengshui People's Hospital,Hengshui 053000)
出处
《现代妇产科进展》
北大核心
2023年第12期924-929,共6页
Progress in Obstetrics and Gynecology
基金
河北省中医药管理局科研计划项目(No:2023287)。
