摘要
目的:探讨那可丁(Nos)对结肠癌SW480细胞钙黏素17(CDH17)表达的影响及其对细胞迁移的作用机制。方法:选用SW480细胞,分为对照组、空载组(si-EV)、CDH17干扰组(si-CDH17)、Nos处理组和CDH17干扰+Nos处理组(si-CDH17+Nos),其中CDH17干扰采用小干扰RNA(siRNA)敲低CDH17水平,Nos处理使用半数抑制质量浓度(55.30±2.21)μg/ml。用qPCR检测SW480细胞CDH17 mRNA表达水平,用Hoechst33258染色法和Transwell实验检测细胞的凋亡和迁移能力,用ELISA检测细胞中VEGF、MMP2和MMP9含量,用WB检测细胞CDH17、Wnt3a和β-catenin蛋白表达水平。结果:与对照组相比,Nos组、si-CDH17组和si-CDH17+Nos组的CDH17 mRNA及蛋白表达水平明显降低(均P<0.01),细胞凋亡增加、迁移能力减弱,细胞中VEGF、MPP2和MPP9含量及Wnt3a和β-catenin蛋白表达水平显著降低(均P<0.01),且si-CDH17+Nos组比si-CDH17组效果更显著(P<0.01)。结论:Nos可诱导人结肠癌SW480细胞凋亡、抑制其迁移能力,其机制可能与下调CDH17表达抑制Wnt3a/β-catenin信号通路有关。
Objective:To explore the effects of noscapine(Nos)on the expression of cadherin 17(CDH17)in colon cancer SW480 cells and the mechanism of Nos on cell migration.Methods:SW480 cells were divided into the control group,empty vector(si-EV)group,CDH17 interference(si-CDH17)group,Nos treatment group,and CDH17 interference+Nos treatment(si-CDH17+Nos)group.Small interfering RNA(siRNA)was used to knockdown CDH17,and the selected concentration of Nos was(55.30±2.21)μg/ml(IC50).The mRNA expression of CDH17 was detected by qPCR;the apoptosis and migration abilities of SW480 cells were observed by Hoechst33258 staining and Transwell assay;the contents of VEGF,MMP2 and MMP9 in SW480 cells were measured by ELISA,and the protein expressions of CDH17,Wnt3a andβ-catenin were determined by WB.Results:Compared with the control group,mRNA and protein expressions of CDH17 obviously decreased,cell apoptosis and migration significantly reduced,while the contents of VEGF,MMP2 and MMP9 as well as the protein expressions of Wnt3a andβ-catenin significantly decreased in Nos treatment group,si-CDH17 group and si-CDH17+Nos treatment group(all P<0.01).The effect of si-CDH17+Nos treatment was more significant than that of si-CDH17(P<0.01).Conclusion:Nos induces apoptosis and inhibits the migration of human colon cancer SW480 cells,which may be related to the down-regulation of CDH17 expression and inhibition of the Wnt3a/β-catenin signaling pathway.
作者
韩峥
黄晓东
刘蒙
朱庆曦
谭洁
刘维洁
陈巍
邹艳丽
蔡一珊
黄莎莎
田霞
HAN Zheng;HUANG Xiaodong;LIU Meng;ZHU Qingxi;TAN Jie;LIUWeijie;CHENWei;ZOUYanli;CAI Yishan;HUANG Shasha;TIAN Xia(Department of Gastroenterology,the Third Hospital ofWuhan City,Wuhan 430060,Hubei,China)
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2019年第10期1089-1094,共6页
Chinese Journal of Cancer Biotherapy
基金
湖北省自然科学基金资助项目（No.2018CFB725）
武汉市卫计委重点项目（No.WX17B05）~~
