虎杖苷通过Hippo/YAP通路影响甲状腺癌8505C细胞的恶性生物学行为和顺铂敏感性  

Polydatin affects malignant biological behaviors and DDP chemosensitivity of thyroid cancer 8505C cells through the Hippo/YAP pathway

作　　者：曹建中[1] 黄金石[1] 丁亚亭[1] CAO Jianzhong;HUANG Jinshi;DING Yating(Department of Pharmacy,Affiliated Hospital of Nantong University,Nantong 226001,Jiangsu,China)

机构地区：[1]南通大学附属医院药学部,江苏南通226001

出　　处：《中国肿瘤生物治疗杂志》2024年第3期224-230,共7页Chinese Journal of Cancer Biotherapy

基　　金：南通市市级科技计划(No.JCZ20004)。

摘　　要：目的:探究虎杖苷通过Hippo/Yes相关蛋白(YAP)通路对人甲状腺癌8505C细胞的恶性生物学行为和顺铂(DDP)敏感性的影响。方法:体外培养8505C细胞,构建其DDP耐药细胞8505C/DDP,用CCK-8法检测0、25、50、75、100 nmol/L虎杖苷处理8505C和8505C/DDP细胞的增殖能力,以筛选虎杖苷的最佳作用浓度。将8505C细胞分为对照组、虎杖苷组、空载组、虎杖苷+YAP1过表达组;将8505C/DDP细胞分为对照组、DDP组、DDP+虎杖苷组、DDP+空载组、DDP+虎杖苷+YAP1过表达组。WB法检测各组8505C细胞中Hippo/YAP通路[YAP1、转录辅激活因子(TAZ)]和EMT(E-cadherin、N-cadherin)相关蛋白,8505C/DDP细胞中YAP1、TAZ、耐药相关蛋白[P-糖蛋白(P-gp)、多药耐药相关蛋白1(MRP1)]、凋亡相关蛋白(C-caspase-3、BAX、Bcl-2)的表达。Transwell小室和细胞划痕实验分别检测各组8505C、8505C/DDP细胞的侵袭、迁移能力。结果:虎杖苷可显著抑制8505C细胞的增殖活性(P<0.05)明显抑制8505C细胞中YAP1、TAZ蛋白、N-cadherin的表达(均P<0.05),提升E-caderin蛋白的表达(P<0.05),显著抑制8505C细胞的迁移和侵袭能力(均P<0.05),而8505C/DDP细胞对低浓度的虎杖苷具有耐药性(P<0.05);过表达YAP1则可逆转虎杖苷对8505C细胞的影响。50 nmol/L虎杖苷明显抑制DDP处理的8505C/DDP细胞中YAP1、TAZ、P-gp、MRP1、Bcl-2的蛋白的表达(均P<0.05),提升cleaved caspase-3、BAX蛋白的表达(均P<0.05)并诱导其细胞凋亡(P<0.05),过表达YAP1则可逆转虎杖苷对8505C/DDP细胞的影响。结论:虎杖苷抑制Hippo/YAP信号通路,从而抑制8505C细胞的恶性生物学行为和增强其对的DDP敏感性。Objective:To investigate the effect of polydatin on malignant biological behaviors and cisplatin(DDP)-sensitivity of human thyroid cancer 8505C cells by regulating the Hippo/Yes-associated protein(YAP)signaling pathway.Methods:8505C cells were cultured in vitro,and their DDP-resistant cells(8505C/DDP)were constructed.The proliferation ability of 8505C and 8505C/DDP cells that treated with 0,25,50,75,and 100 nmol/L polydatin was detected by CCK-8 assay,in order to screen the optimal action concentration of polydatin.The 8505C cells were divided into control group,polydatin group,empty vector group,polydatin+YAP1 overexpression group;and the 8505C/DDP cells were divided into control group,DDP group,DDP+polydatin group,DDP+empty vector group,and DDP+polydatin+YAP1 overexpression group.WB assay was used to detect the expression of Hippo/YAP pathway related proteins[YAP1,transcriptional coactivator factor(TAZ)]and EMT-associated proteins(E-cadherin,N-cadherin)in the 8505C cells of each group;Besides,the expression levels of YAP1,TAZ,and drug resistance-associated proteins[P-glycoprotein(P-gp),multidrug resistance-associated protein 1(MRP1)]as well as apoptosis-associated proteins(cleaved caspase-3,BAX,Bcl-2)in 8505C/DDP cells were also detected by WB.Transwell assay and cell scratching assay were used to detect the invasion and migration abilities of 8505C and 8505C/DDP cells in each group,respectively.Results:Polydatin significantly inhibited the proliferation of 8505C cells(P<0.05),but 8505C/DDP cells were resistant to low concentrations of polydatin(P<0.05).Moreover,polydatin significantly inhibited the protein expression of YAP1,TAZ and N-cadherin,elevated the protein expression of E-cadherin in 8505C cells(P<0.05),and significantly suppressed the migration and invasion of 8505C cells(all P<0.05);However,overexpression of YAP1 reversed the effects of polydatin on 8505C cells(all P<0.05).50 nmo/L polydatin significantly inhibited the protein expression of YAP1,TAZ,P-gp,MRP1 and Bcl-2,elevated the protein expression

关 键 词：甲状腺癌 8505C细胞 虎杖苷 Hippo/YAP通路 迁移 侵袭 上皮间质转化 化疗敏感性 

分 类 号：R736.1[医药卫生—肿瘤] R730.5[医药卫生—临床医学]