毛钩藤碱对人宫颈癌Ca Ski细胞增殖、凋亡、转移及侵袭的影响  

Effect of Hirsutine on Proliferation,Apoptosis,Metastasis,and Invasion of Human Cervical Cancer Ca Ski Cells

作　　者：李亚威[1] 谢磊[1] 王娜[1] LI Ya-wei;XIE Lei;WANG Na(The First Hospital of Hebei Medical University,Shijiazhuang 050000,China)

机构地区：[1]河北医科大学第一医院,石家庄050000

出　　处：《中国实验方剂学杂志》2022年第4期109-115,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：河北省医学科学研究课题项目(20210081)。

摘　　要：目的:探讨毛钩藤碱对人宫颈癌Ca Ski细胞增殖、凋亡、转移、侵袭的影响及机制。方法:细胞活性与增殖检测(CCK-8)法检测细胞增殖活性,Hoechst 33258染色及流式细胞术检测细胞凋亡,实时荧光定量聚合酶链式反应检测B细胞淋巴瘤-2(Bcl-2),Bcl-2相关X蛋白(Bax),p53 mRNA表达,划痕实验检测细胞转移能力,Transwell小室实验检测细胞侵袭能力,酶联免疫吸附测定(ELISA)检测纤连蛋白(FN),基质金属蛋白酶-2(MMP-2)及基质金属蛋白酶-9(MMP-9)含量,蛋白免疫印迹法(Western blot)检测蛋白表达。结果:与空白组比较,毛钩藤碱可以呈剂量及时间依赖性的抑制Ca Ski细胞的增殖;8.0,16.0,32.0μmol·L^(-1)毛钩藤碱组细胞凋亡率,Bax,p53 mRNA表达增加(P<0.05,P<0.01),Bcl-2 mRNA及磷酸化Src(p-Src),磷酸化信号转导和转录活化因子3(p-STAT3)蛋白表达降低(P<0.05,P<0.01)。与空白组比较,1.0,2.0,4.0μmol·L^(-1)毛钩藤碱组细胞迁移率,穿膜细胞数,FN,MMP-2,MMP-9含量及缺氧诱导因子-1α(HIF-1α),波形蛋白(Vimentin),N-钙黏蛋白(N-cadherin)表达降低(P<0.05,P<0.01),E-钙黏蛋白(E-cadherin)表达增加(P<0.01)。结论:毛钩藤碱可以抑制人宫颈癌Ca Ski细胞的增殖、转移及侵袭,并能诱导凋亡,作用机制与调控Src/STAT3及HIF-1α/上皮间充质转化(EMT)信号通路有关。Objective:To investigate the effects of hirsutine on proliferation,apoptosis,metastasis,and invasion of human cervical cancer Ca Ski cells and its action mechanism.Method:The cell proliferation was determined by cell counting kit-8(CCK-8)assay and the cell apoptosis by Hoechst 33258 staining and flow cytometry.The mRNA expression levels of B cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax),and p53 were detected by real-time fluorescence quantitative polymerase chain reaction.The scratch test was conducted to detect the cell mobility,followed by the detection of cell invasion ability using a Transwell chamber.The contents of fibronectin(FN),matrix metalloproteinase-2(MMP-2),and matrix metalloproteinase-9(MMP-9)were measured by enzyme linked immunosorbent assay(ELISA),and their protein expression levels were assayed by Western blot.Result:Compared with the control group,hirsutine inhibited the proliferation of Ca Ski cells in a dose-and time-dependent manner.The apoptosis rates and Bax and p53 mRNA expression levels in the 8.0,16.0,32.0μmol·L^(-1) hirsutine groups rose(P<0.05,P<0.01),while the Bcl-2 mRNA expression and phosphorylated Src(p-Src)and phosphorylated signal transducer and activator of transcription factor 3(STAT3)protein expression declined(P<0.05,P<0.01).Compared with the control group,the 1.0,2.0,4.0μmol·L^(-1) hirsutine groups exhibited lowered cell mobility,number of transmembrane cells,FN,MMP-2 and MMP-9 contents,and hypoxia inducible factor-1α(HIF-1α),Vimentin,and N-cadherin protein expression(P<0.05,P<0.01),but elevated E-cadherin protein expression(P<0.01).Conclusion:Hirsutine inhibits the proliferation,metastasis,and invasion of human cervical cancer Ca Ski cells and induces their apoptosis,which may be related to its regulation of Src/STAT3 and HIF-1α/epithelial mesenchymal transformation(EMT)signaling pathways.

关 键 词：毛钩藤碱 宫颈癌 增殖 凋亡 转移 侵袭 

分 类 号：R22[医药卫生—中医基础理论] R242[医药卫生—中医学]