蔓荆子黄素调控miR-378/PRRX1轴对胃癌生物学行为的影响  

Impacts of Casticin on biological behavior of gastric cancer cells by regulating miR-378/PRRX1 axis

作　　者：张丛 冯华 黄小龙 王旋 ZHANG Cong;FENG Hua;HUANG Xiaolong;WANG Xuan(Department of General Surgery,Xingtai People's Hospital,Xingtai 054000,China)

机构地区：[1]邢台市人民医院普外科,054000

出　　处：《天津医药》2023年第10期1040-1047,共8页Tianjin Medical Journal

基　　金：河北省医学科学研究课题计划项目(20201574)。

摘　　要：目的探究蔓荆子黄素(Cas)对胃癌(GC)SGC-7901细胞增殖、侵袭、迁移及微小RNA-378(miR-378)/配对相关同源框1(PRRX1)轴的影响。方法不同浓度Cas(0、5、10、20、40、80μmol/L)处理SGC-7901细胞48 h。将对数生长期的SGC-7901细胞分为:对照组、Cas低浓度组(10μmol/L)、Cas中浓度组(20μmol/L)、Cas高浓度组(40μmol/L)、Cas高浓度+miR-378小干扰RNA(siRNA)组(40μmol/L Cas+miR-378 siRNA)、Cas高浓度+PRRX1组(40μmol/L Cas+PRRX1过表达质粒)、Cas高浓度+miR-378 siRNA+PRRX1组(40μmol/L Cas+miR-378 siRNA+PRRX1过表达质粒)。MTT法、集落形成实验、流式细胞术、Transwell小室、划痕实验分别测定各组SGC-7901细胞活力、集落形成数量、凋亡率、侵袭及迁移能力;qPCR、蛋白印迹实验分别检测SGC-7901细胞miR-378、PRRX1蛋白表达水平;双萤光素酶实验验证miR-378和PRRX1靶向关系;体内异种移植SGC-7901细胞建立裸鼠移植瘤模型,验证Cas的抗癌活性及对miR-378表达的影响。结果Cas 10、20、40、80μmol/L组SGC-7901细胞活力呈浓度依赖性降低(P<0.05),选取10、20、40μmol/L的Cas作为后续研究浓度。与对照组相比,Cas低、中、高浓度组SGC-7901细胞活力、集落形成数量、侵袭细胞数量、划痕愈合率、PRRX1蛋白表达水平依次降低(P<0.05),细胞凋亡率和miR-378表达水平依次升高(P<0.05);与Cas高浓度组相比,Cas高浓度+miR-378 siRNA组和Cas高浓度+PRRX1组SGC-7901细胞活力、集落形成数量、侵袭细胞数量、划痕愈合率、PRRX1蛋白表达水平升高,细胞凋亡率降低(P<0.05),Cas高浓度+miR-378 siRNA组SGC-7901细胞miR-378表达水平降低(P<0.05);与Cas高浓度+miR-378 siRNA组相比,Cas高浓度+PRRX1组SGC-7901细胞miR-378表达水平升高(P<0.05);与Cas高浓度+miR-378 siRNA组、Cas高浓度+PRRX1组相比,Cas高浓度+miR-378 siRNA+PRRX1组SGC-7901细胞活力、集落形成数量、侵袭细胞数量、划痕愈合率、PRRX1蛋白表达水平升高(P<0.05),细胞凋亡率�Objective To explore effects of Casticin(Cas)on the proliferation,invasion,migration and microRNA-378(miR-378)/paired related homeobox 1(PRRX1)axis of gastric cancer(GC)SGC-7901 cells.Methods SGC-7901 cells were treated with Cas at different concentrations(0,5,10,20,40 and 80μmol/L)for 48 h,and the cell viability of SGC 7901 was detected by tetrazole salt(MTT)method.SGC-7901 cells at logarithmic growth stage were divided into 7 groups:the control group,the Cas low concentration group(10μmol/L),the Cas medium concentration group(20μmol/L),the Cas high concentration group(40μmol/L),the Cas high concentration+miR-378 small interfering RNA(siRNA)group(40μmol/L Cas+miR-378 siRNA),the Cas high concentration+PRRX1 group(40μmol/L Cas+PRRX1 overexpressed plasmid)and the Cas high concentration+miR-378 siRNA+PRRX1 group(40μmol/L Cas+miR-378 siRNA+PRRX1 overexpression plasmid).Cell activity,colony formation number,apoptosis rate,invasion and migration ability of SGC-7901 cells were determined by MTT assay,colony formation assay,flow cytometry,Transwell chamber assay and scratch assay,respectively.Expression levels of miR-378 and PRRX1 protein in SGC-7901 cells were detected by fluorescence quantitative PCR and Western blot assay.Double luciferase assay verified the targeting relationship between miR-378 and PRRX1.Xenografted SGC-7901 cells were xenografted into nude mice to verify the anticancer activity of Cas and its effect on the expression of miR-378.Results The cell viability of SGC-7901 cells in Cas 10,20,40 and 80μmol/L groups decreased in a concentration-dependent manner(P<0.05),and 10,20 and 40μmol/L Cas were selected as the follow-up study concentration.Compared with the control group,the cell viability,colony formation number,invasion cell number,scratch healing rate and PRRX1 protein expression level of SGC-7901 cells in the Cas low,medium and high concentration groups were decreased successively(P<0.05),while the apoptosis rate and the expression level of miR-378 were increased successively(P<0.05).Comp

关 键 词：蔓荆子 胃肿瘤 微RNAS 细胞增殖 细胞运动 肿瘤浸润 同源盒结构域蛋白质类 

分 类 号：R285.5[医药卫生—中药学] R735.2[医药卫生—中医学]