益气解毒方通过TGF-β1/SMAD3信号通路抑制鼻咽癌细胞增殖、迁移和侵袭  

Experimental Study on Inhibitory Effect of Yiqi Jiedu Recipe on Proliferation,Migration and Invasion of Nasopharyngeal Carcinoma Cells Through TGF-β1/SMAD3 Signaling Pathway

作　　者：郭利培 张文青 刘洁[1] 史红健[1,2] 何迎春 王贤文[1,2] 范婧莹 GUO Lipei;ZHANG Wenqing;LIU Jie;SHI Hongjian;HE Yingchun;WANG Xianwen;FAN Jingying(Hunan University of Chinese Medicine,Changsha 410208 Hunan,China;Hunan Provincial Key Laboratory for the Prevention and Treatment of Ophthalmology and Otolaryngology Diseases with Traditional Chinese Medicine,Changsha 410208 Hunan,China)

机构地区：[1]湖南中医药大学,湖南长沙410208 [2]中医药防治眼耳鼻咽喉疾病湖南省重点实验室,湖南长沙410208

出　　处：《中药新药与临床药理》2024年第7期935-943,共9页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金项目(81973914);湖南省教育厅科学研究项目(21B0358,22A0254);湖南省中医药管理局资助项目(D2022105);2021年度湖南中医药大学校级科研基金(2021XJJJ014,2021XJJJ008);湖南省卫健委项目(D202307017740)。

摘　　要：目的研究益气解毒方对鼻咽癌细胞增殖、迁移和侵袭的影响,并从转化生长因子β1(TGF-β1)/SMAD3信号通路探讨其对增殖、迁移和侵袭的作用机制。方法(1)将鼻咽癌细胞分为4组:溶剂对照组、益气解毒方0.5 mg·mL^(-1)组、益气解毒方1.0 mg·mL^(-1)组、5-氟尿嘧啶(5-Fluorouracil,5-Fu)2μg·mL^(-1)组,采用实时无标记细胞功能分析仪(real time cellular analysis technology,RTCA)监测细胞增殖;创伤愈合实验检测细胞迁移;药物干预24 h后,侵袭小室法检测细胞侵袭;Western Blot法检测细胞β-catenin、E-cadherin、N-cadherin、TGF-β1、SMAD3蛋白的表达水平。(2)将鼻咽癌细胞分为5组:溶剂对照组、TGF-β110 ng·mL^(-1)组、TGF-β110 ng·mL^(-1)+益气解毒方1.0 mg·mL^(-1)组、益气解毒方1.0 mg·mL^(-1)组、LY320088210μmol·L-1组,采用实时无标记细胞功能分析仪监测细胞增殖;创伤愈合实验检测细胞迁移;药物干预24 h后,侵袭小室法检测细胞侵袭;Western Blot法检测细胞β-catenin、E-cadherin、N-cadherin、TGF-β1、SMAD3蛋白的表达水平。(3)随机将裸鼠分为模型组、益气解毒方组和5-Fu组。皮下部位注射细胞悬液制备鼻咽癌裸鼠移植瘤模型,基本成瘤后,5-Fu组每2 d腹腔注射1次,其余组每天灌胃给药1次,连续3周。每隔3 d测量瘤体体积1次;Western Blot法检测各组组织中β-catenin、E-cadherin、N-cadherin蛋白表达水平。结果与溶剂对照组比较,益气解毒方(0.5、1.0 mg·mL^(-1))组的细胞增殖曲线均下降,细胞迁移和侵袭能力均降低(P<0.05,P<0.01),且E-cadherin蛋白表达明显升高(P<0.01),β-catenin、N-cadherin、TGF-β1、SMAD3蛋白表达均明显降低(P<0.05,P<0.01)。与模型组比较,益气解毒方组的鼻咽癌移植瘤体积明显减小(P<0.05),且益气解毒方组移植瘤组织中的β-catenin和N-cadherin蛋白表达明显下调(P<0.05,P<0.01),E-cadherin蛋白表达明显上调(P<0.01)。加入TGF-β1信号通路激活剂和抑制剂后,与�Objective To investigate the effect of Yiqi Jiedu Recipe(YQ)on the proliferation,migration,and invasion of nasopharyngeal carcinoma cells,and to explore its mechanisms of action on proliferation,migration,and invasion through the TGF-β1/SMAD3 signaling pathway.Methods(1)The 5-8F cells were divided into four groups:solvent control group,YQ 0.5 mg·mL^(-1)group,YQ 1.0 mg·mL^(-1)group,and 5-fluorouracil 2μg·mL^(-1)group.Cell proliferation was monitored using real-time cell analysis(RTCA).Wound healing experiment was conducted to assess cell migration.After 24 hours of drug intervention,transwell assay was employed to measure cell invasion.The protein expression levels ofβ-catenin,E-cadherin,N-cadherin,TGF-β1,and SMAD3 in the cells were evaluated using the Western Blot method.(2)The 5-8F cells were divided into five groups:solvent control group,TGF-β110 ng·mL^(-1)group,TGF-β110 ng·mL^(-1)+YQ 1.0 mg·mL^(-1)group,YQ 1.0 mg·mL^(-1)group,and LY320088210μmol·L-1 group.Cell proliferation was monitored using RTCA.Wound healing experiment was conducted to assess cell migration.After 24 hours of drug intervention,transwell assay was employed to measure cell invasion.The protein expression levels ofβ-catenin,E-cadherin,N-cadherin,TGF-β1,and SMAD3 in the cells were evaluated using Western Blot.(3)The nude mice were randomly assigned into the model group,YQ group,and 5-fluorouracil group.Subcutaneous injection of 5-8F cell suspension was performed to establish the xenograft nude mouse model of nasopharyngeal carcinoma.After the tumors reached a certain size,the 5-fluorouracil group received intraperitoneal injection of 5-Fu once every 2 days,while the other groups were orally administered corresponding drugs once a day for three consecutive weeks.Tumor volume was measured every 3 days.Western Blot was conducted to assess the protein expression levels ofβ-catenin,E-cadherin,and N-cadherin in the tissues of each group.Results Compared with the solvent control group,the proliferation curves of 5-8F cells in the YQ(

关 键 词：益气解毒方 鼻咽癌 TGF-β1/SMAD3信号通路 增殖 迁移 侵袭 BALB/c-nu裸鼠 人高转移鼻咽癌5-8F细胞 

分 类 号：R285.5[医药卫生—中药学]