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 of cleaved caspase-3 and BAX in 8505C/DDP cells which treated with DDP (all P<0.05), and induced cell apoptosis (all P<0.05); However, overexpression of YAP1 reversed the effect of polydatin on 8505C/DDP cells (all P<0.05). Conclusion: Polydatin inhibits the malignant biological behaviors and enhances the DDP-sensitivity of 8505C cells via inhibiting the Hippo/YAP signaling pathway.

南通市市级科技计划（No.JCZ20004）