WWOX对胆囊癌细胞迁移、侵袭的调控作用及机制  

Regulation and mechanism of WWOX on migration and invasion of gallbladder cancer cells

作　　者：陈广 魏东[1] 苏琨[1] 雷学芬 王滔[1] 洪茂林 房立锐 CHEN Guang;WEI Dong;SU Kun;LEI Xuefen;WANG Tao;HONG Maolin;FANG Lirui(Department of Hepatobiliary Surgery,the Second Affiliated Hospital of Kunming Medical University,Kunming 650000,China;不详)

机构地区：[1]昆明医科大学第二附属医院肝胆胰外科,昆明650000 [2]昆明医科大学第二附属医院肿瘤科

出　　处：《山东医药》2021年第19期20-24,共5页Shandong Medical Journal

基　　金：云南省应用基础研究基金资助项目[2019FE001(-071)];昆明医科大学第二附属医院院内科技计划项目(2018yk016)。

摘　　要：目的探讨含WW域的氧化还原酶(WWOX)对胆囊癌细胞迁移、侵袭的调控作用及机制。方法培养胆囊癌细胞(GBC-SD),WWOX过表达的慢病毒质粒和包装质粒共转染293T细胞获得慢病毒颗粒。将GBC-SD细胞随机分为三组,分别加入等量的WWOX过表达的慢病毒(WWOX组)、阴性对照慢病毒(NEG组)、完全培养液(GBC-SD组)。用qRT-PCR检测细胞内WWOX、TGF-β_(1) mRNA表达,用Western blotting法检测细胞内WWOX、TGF-β_(1)蛋白表达,用划痕实验检测细胞迁移能力,用Transwell侵袭实验检测细胞侵袭能力,用ELISA法检测GBC-SD细胞上清液中TGF-β_(1)。结果与NEG组、GBC-SD组比较,WWOX组细胞WWOX mRNA及蛋白相对表达量较高(P均<0.05);NEG组、GBC-SD组细胞WWOX mRNA及蛋白相对表达量比较差异均无统计学意义(P均>0.05)。与NEG组、GBC-SD组比较,WWOX组细胞划痕愈合度低、细胞侵袭数少(P均<0.05),NEG组、GBC-SD组细胞划痕愈合度、细胞侵袭数比较差异均无统计学意义(P均>0.05)。与NEG组、GBC-SD组比较,WWOX组细胞内TGF-β_(1) mRNA、蛋白相对表达量低,细胞上清液中TGF-β_(1)水平低(P均<0.05);NEG组、GBC-SD组细胞内TGF-β_(1) mRNA、蛋白相对表达量,细胞上清液中TGF-β_(1)水平比较差异均无统计学意义(P均>0.05)。结论WWOX过表达可抑制GBC-SD细胞迁移、侵袭,其机制可能与抑制TGF-β_(1)的表达及胞外分泌有关。Objective To explore the regulation and mechanism of WW-domain-containing oxidoreductase(WWOX)on the migration and invasion of gallbladder cancer cells.Methods Gallbladder cancer cells(GBC-SD)were cultured,and WWOX overexpressed lentiviral plasmid and packaging plasmid were co-transfected into 293T cells to obtain lentiviral particles.The GBC-SD cells were randomly divided into three groups,which were added with the same amount of WWOX overexpressing lentivirus(WWOX group),negative control lentivirus(NEG group),and complete culture medium(GBC-SD group),respectively.WWOX and transforming growth factor-β_(1)(TGF-β_(1))mRNA in cells were detected by qRT-PCR,and WWOX and TGF-β_(1) protein in cells were detected by Western blotting.Scratch test was used to detect the cell migration ability,Transwell invasion test was used to detect the cell invasion ability,and ELISA was used to detect TGF-β_(1) in the culture supernatant of GBC-SD cells.Results Compared with the NEG group and GBC-SD group,the relative expression of WWOX mRNA and protein in the WWOX group was higher(P<0.05).There were no significantly differences in the relative expression of WWOX mRNA and protein between NEG group and GBC-SD group(P>0.05).Compared with the NEG group and GBC-SD group,the WWOX group had lower cell wound healing and fewer invasion cells(all P<0.05).No significant differences were found in the cell scratch healing degree or cell invasion number between the NEG group and GBC-SD group(all P>0.05).Compared with the NEG group and GBC-SD group,the relative expression of TGF-β_(1) mRNA and protein in the WWOX group was lower,and the level of TGF-β_(1) in cell supernatant was lower(P<0.05).There were no significant differences in the relative expression of TGF-β_(1) mRNA and protein or the level of TGF-β_(1) in the cell supernatant between NEG group and GBC-SD group(all P>0.05).Conclusion Overexpression of WWOX can significantly inhibit the migration and invasion of GBC-SD cells,which may be achieved by inhibiting the expression and ext

关 键 词：胆囊癌 WW域的氧化还原酶 转化生长因子β_(1) 细胞迁移 细胞侵袭 

分 类 号：R735.8[医药卫生—肿瘤]