益气解毒方通过Wnt/β-catenin信号通路对鼻咽癌CNE2细胞增殖的影响  

Effects of Yiqi Jiedu Formula on Proliferation of Nasopharyngeal Carcinoma CNE2 Cells by Wnt/β-catenin Signaling Pathway

作　　者：范婧莹[1,2] 刘洁 刘晓丹[1] 戴娜 何兰[1,2] 王贤文 李丽鹏[1] 邹韵 何迎春 FAN Jingying;LIU Jie;LIU Xiaodan;DAI Na;HE Lan;WANG Xianwen;LI Lipeng;ZOU Yun;HE Yingchun(Hunan University of Chinese Medicine,Changsha,Hunan 410208,China;Hunan Provincial Key Laboratory for the Prevention and Treatment of Ophthalmology and Otolaryngology Diseases with Traditional Chinese Medicine,Changsha,Hunan 410208,China;The First Affiliated of Hunan University of Chinese Medicine,Changsha,Hunan 410007,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 [3]湖南中医药大学第一附属医院,湖南长沙410007

出　　处：《湖南中医药大学学报》2020年第5期535-539,共5页Journal of Hunan University of Chinese Medicine

基　　金：国家自然科学基金资助项目(81573721,81874408,81973914);湖南省自然科学基金(2019JJ40216);湖南省教育厅科研基金项目(16C1212);2018年湖南中医药大学校级大学生研究性学习和创新性实验计划课题(2018-072)。

摘　　要：目的探讨益气解毒方对鼻咽癌CNE2细胞增殖及Wnt/β-catenin信号通路的影响。方法将CNE2细胞用不同浓度(0.25、0.50、1.0 g/L)的益气解毒方水提物和阳性对照药(顺铂,4 mg/L)分别处理24、48、72 h,采用MTT法检测细胞增殖率,流式细胞术检测药物作用48 h后细胞周期分布情况;Western blot检测Wnt/β-catenin信号通路关键蛋白Wnt5a/b、β-catenin及细胞周期蛋白CyclinD1的表达情况。结果MTT结果显示,益气解毒方对鼻咽癌CNE2细胞增殖具有明显抑制作用,且其抑制作用与作用时间、药物浓度呈正相关(P<0.05)。细胞周期结果显示,处理48 h后,G0/G1期比例降低,S期和G2/M期比例增加(P<0.05,P<0.01)。Western blot结果显示,水提物处理48 h后,Wnt/β-catenin信号通路关键蛋白Wnt5a/b、β-catenin及细胞周期蛋白CyclinD1的表达量明显下降(P<0.01)。结论益气解毒方可通过下调Wnt/β-catenin信号通路关键蛋白Wnt5a/b、β-catenin及细胞周期蛋白CyclinD1的表达,阻滞细胞周期,抑制人鼻咽癌CNE2细胞的增殖。Objective To explore the effects of Yiqi Jiedu Formula(YQ)on proliferation of CNE2 cells and Wnt/β-catenin signaling pathway in nasopharyngeal carcinoma.Methods CNE2 cells were respectively treated with different concentrations(0.25,0.50,1.0 g/L)of YQ water extract and positive control medicine(cisplatin,4 mg/L)for 24,48,and 72 h.Then cell proliferation rate activity was detected by MTT method and the cell cycle distribution after 48 h was detected by flow cytometry.Western blot method was used to detect the expression levels of key proteins Wnt5a/b,β-catenin and CyclinD1 in Wnt/β-catenin signaling pathway.Results MTT results showed that YQ group had significant inhibitory effect on the proliferation of nasopharyngeal carcinoma CNE2 cells,and the inhibitory effect was positively correlated with the time and medicine concentration(P<0.05).The results of cell cycle showed that after 48 h of treatment,the proportion of G0/G1 was decreased,and the proportion of S and G2/M was increased(P<0.05,P<0.01).Western blot results showed that after 48 h of water extraction treatment,the expression levels of Wnt/β-catenin signaling pathway related protein Wnt5a/b,β-catenin and CyclinD1 decreased significantly(P<0.01).Conclusion YQ could inhibit the proliferation of human nasopharyngeal carcinoma CNE2 cells and block cell cycle by down-regulating the expression of Wnt/β-catenin signaling pathway key proteins Wnt5a/b,β-catenin and CyclinD1.

关 键 词：益气解毒方 鼻咽癌 WNT/Β-CATENIN信号通路 细胞周期 细胞增殖 

分 类 号：R285.5[医药卫生—中药学]