基于Hippo/yap/TAZ途径探讨雷公藤内酯醇-聚乙烯亚胺-环糊精逆转卵巢癌耐药的作用及机制  

Effect and Mechanism of Triptolide-polyethleneimine-cyclodextrin on Reversing Drug Resistance of Ovarian Cancer through Hippo/yap/TAZ Signaling Pathway

作　　者：黄根华[1] 谭布珍[1] 占伏良[1] 胡辉[1] HUANG Genhua;TAN Buzhen;ZHAN Fuliang;HU Hui(The Second Affiliated Hospital of Nanchang University,Nanchang 330006 Jiangxi,China)

机构地区：[1]南昌大学第二附属医院

出　　处：《中药新药与临床药理》2020年第2期185-191,共7页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：江西省卫生计生委科技计划项目（20195243）

摘　　要：目的观察雷公藤内酯醇-聚乙烯亚胺-环糊精复合物(TP-PEI-CyD)对人卵巢癌顺铂耐药细胞(SKOV3/DDP)的作用及其可能的作用机制。方法采用CCK-8法测定雷公藤内酯醇单体(triptolide,TPL)、TP-PEICyD对人正常卵巢上皮细胞IOSE80、卵巢癌耐药细胞SKOV3/DDP的细胞毒性,筛选TP-PEI-CyD对SKOV3/DDP的低毒浓度;采用低毒浓度TP-PEI-CyD、TPL处理SKOV3/DDP细胞,流式细胞术检测SKOV3/DDP细胞凋亡,划痕实验检测SKOV3/DDP细胞迁移,Transwell检测SKOV3/DDP细胞侵袭,RT-PCR及Western Blot法检测SKOV3/DDP细胞内Hippo、yap及TAZ mRNA和蛋白表达。结果与对照组比较,TPL、TP-PEICyD均可明显抑制IOSE80、SKOV3/DDP细胞的生长,且细胞毒性随着TPL、TP-PEI-CyD浓度的升高而增加,与TPL组相比,TP-PEI-CyD对IOSE80细胞的毒性明显降低(P<0.05),但对SKOV3/DDP细胞的抑制率无明显变化;低毒浓度的TP-PEI-CyD、TPL作用于SKOV3/DDP细胞后:TP-PEI-CyD组、TPL组SKOV3/DDP细胞凋亡率显著增加(P<0.01);迁移能力、侵袭能力显著降低(P<0.01);SKOV3/DDP细胞中Hippo、yap及TAZ的mRNA及蛋白表达均显著降低(P<0.05);而TP-PEI-CyD组与TPL单体组相比,差异均无统计学意义(P>0.05);结论雷公藤内酯醇耦合聚乙烯亚胺-环糊精后,其毒副作用降低,且可有效逆转SKOV-3细胞的肿瘤耐药性,其疗效等同于雷公藤内酯醇单药,其可能是通过调节Hippo/yap/TAZ信号通路来实现的。Objective To study the effect of triptolide-polyethleneimine-cyclodextrin(TP-PEI-CyD) complex on SKOV3/DDP cells and its underlying mechanism.Methods The growth inhibitory effects of triptolide(TPL),TPPEI-CyD on IOSE80,SKOV3/DDP cells were examined by CCK-8 assay.The low-toxic concentration of TP-PEICyD on SKOV3/DDP cells were calculated.Then SKOV3/DDP cells were treated with low-toxic concentration of TPPEI-CyD and TPL respectively.The apoptosis of SKOV3/DDP cells was detected by flow cytometry.The cell migration of SKOV3/DDP was detected by scratch assay.The cell invasion of SKOV3/DDP was detected by Transwell.The mRNA and protein expression levels of Hippo,yap and TAZ in SKOV3/DDP cells were detected by RT-PCR and Western Blot.Results Compared with the control group,the growth of IOSE80 and SKOV3/DDP cells in TPL and TP-PEI-CyD groups were significantly inhibited,and the cytotoxicity increased with the increasing of drug concentration.Compared with the TPL group,the toxicity of IOSE80 cells in TP-PEI-CyD was significantly reduced(P<0.05),but there was no significant change in the inhibition rate of SKOV3/DDP cells.After SKOV3/DDP cells were treated with low dose TP-PEI-CyD(2 g·mL^-1) and TPL,the apoptosis rates of SKOV3/DDP cells were significantly increased(P<0.01).Cell migration and cell invasion were significantly reduced(P<0.01).mRNA and protein expression levels of Hippo,yap and TAZ in SKOV3/DDP cells were significantly decreased(P<0.05).There was no significant difference between TP-PEI-CyD and TPL groups(P> 0.05).Conclusion After the coupling of tripterygolide with polyethylenei mine cyclodextrin,the toxic effects of TPL were reduced,and the tumor resistance of SKOV3 cells could be effectively reversed,the effect was equal to TPL,which may be achieved by regulating the Hippo/yap/TAZ signaling pathway.

关 键 词：雷公藤内酯醇-聚乙烯亚胺-环糊精复合物 卵巢癌 逆转 耐药 毒副作用 Hippo/yap/TAZ信号通路 

分 类 号：R285.5[医药卫生—中药学]