甜橙黄酮调节YAP/TAZ轴对宫颈癌细胞增殖、凋亡和上皮间质转化的影响  

Effects of sinensetin on the proliferation,apoptosis,and epithelial mesenchymal transition of cervical cancer cells by regulating YAP/TAZ axis

作　　者：谢旭 张欣萍[1] 郭丽萍 许欣 XIE Xu;ZHANG Xinping;GUO Liping;XU Xin(Nanyang First People's Hospital,Henan Nanyang 473000,China.)

机构地区：[1]南阳市第一人民医院,河南南阳473000

出　　处：《现代肿瘤医学》2024年第15期2695-2703,共9页Journal of Modern Oncology

基　　金：江苏省放射医学与防护重点实验室开放课题(编号:KJS2242)。

摘　　要：目的:探究甜橙黄酮(Sinensetin, SIN)调节Yes相关蛋白(YAP)/PDZ结合域的转录共刺激因子(TAZ)轴对宫颈癌细胞增殖、凋亡和上皮间质转化的影响。方法:免疫组织化学和实时荧光定量PCR(qRT-PCR)检测120例宫颈癌组织、癌旁组织中YAP、TAZ表达。以不同浓度SIN(0、5、10、25、50、100、200μg/mL)处理宫颈癌细胞,检测细胞活力。将SiHa细胞分为对照(control)组、SIN低、中、高剂量(SIN-L、M、H)组、SIN-H+XMU-MP-1组,EdU检测细胞增殖,流式细胞术检测细胞凋亡,Transwell小室实验检测细胞迁移和侵袭,Western blot检测YAP、TAZ、PCNA、Bax、Bcl-2、N-cadherin、Vimentin、E-cadherin蛋白表达水平。构建宫颈癌裸鼠模型,分为NC组、SIN组、SIN-H+XMU-MP-1组,测量肿瘤质量与体积,HE染色观察肿瘤组织形态,免疫组化法检测肿瘤组织Ki67、YAP、TAZ蛋白表达。结果:120例宫颈癌组织中YAP阳性率、TAZ阳性率显著升高(P<0.05);宫颈癌组织中YAP、TAZ mRNA表达水平显著高于癌旁组织(P<0.05)。宫颈癌细胞CaSki、C-33A、HeLa、SiHa细胞活力随着SIN浓度的升高而逐渐降低(P<0.05),选择SiHa作为后续实验细胞,选择25、50和100μmol/L的SIN作为后续实验浓度。与对照组相比,SIN-L、M、H组细胞EdU阳性率、迁移细胞数、侵袭细胞数、PCNA、Bcl-2、N-cadherin蛋白表达显著下降,凋亡率、Bax、E-cadherin蛋白表达显著升高(P<0.05);与SIN-H组相比,SIN-H+XMU-MP-1组细胞EdU阳性率、迁移细胞数、侵袭细胞数、PCNA、Bcl-2、N-cadherin蛋白表达显著升高,凋亡率、Bax、E-cadherin蛋白表达显著降低(P<0.05)。SIN能够抑制肿瘤体积和质量,促进肿瘤组织坏死,降低Ki67、YAP、TAZ阳性表达。结论:SIN能够抑制宫颈癌细胞增殖和上皮间质转化,促进凋亡,其作用机制可能与抑制YAP/TAZ轴有关。Objective:To investigate the effects of Sinensetin(SIN)on the proliferation,apoptosis,and epithelial mesenchymal transition of cervical cancer cells by regulating the Yes-associated protein(YAP)/transcriptional coactivator with PDZ-binding motif(TAZ)axis.Methods:Immunohistochemistry and real-time fluorescence quantitative PCR(qRT-PCR)were applied to detect the expression of YAP and TAZ in 120 cervical cancer tissues and adjacent tissues.Cervical cancer cells were treated with different concentrations of SIN(0,5,10,25,50,100,200μg/mL)to detect cell viability.SiHa cells were grouped into control group,SIN low,medium,and high dose(SIN-L,M,H)group,and SIN-H+XMU-MP-1 group,EdU was applied to detect cell proliferation,flow cytometry was applied to detect cell apoptosis,Transwell chamber experiment was applied to detect cell migration and invasion,Western blot was applied to detect the expression levels of YAP,TAZ,PCNA,Bax,Bcl-2,N-cadherin,Vimentin,and E-cadherin proteins.Cervical cancer nude mouse model was constructed and separated into NC group,SIN group,and SIN-H+XMU-MP-1 group,the mass and volume of tumors were measured,HE staining was applied to observe the morphology of tumor tissue,immunohistochemical method was applied to detect the expression of Ki67,YAP and TAZ proteins in tumor tissue. Results: The positive rates of YAP and TAZ were greatly increased in 120 cervical cancer tissues ( P <0.05).The expression levels of YAP and TAZ mRNA in cervical cancer tissue were obviously higher than those in adjacent tissues ( P <0.05).The cell viability of cervical cancer cells CaSki,C-33A,HeLa,and SiHa gradually decreased with the increase of SIN concentration ( P <0.05).SiHa was selected as the subsequent experimental cells,and 25,50,and 100 μmol/L SIN were selected as the subsequent experimental concentrations.Compared with the control group,the positive rate of EdU,the numbers of migrating cells and invasive cells,and the protein expression of PCNA,Bcl-2,and N-cadherin in SIN-L,M,and H groups were obviously reduced,

关 键 词：甜橙黄酮 宫颈癌 增殖 凋亡 上皮间质转化 Yes相关蛋白/PDZ结合域的转录共刺激因子轴 

分 类 号：R737.33[医药卫生—肿瘤]