摘要
目的观察转染过表达WW域的氧化还原酶(WWOX)质粒对胆囊癌细胞株GBC-SD迁移、侵袭能力的变化,并探讨其机制。方法胆囊癌细胞株GBC-SD分为实验组、NEG组、空白对照组,实验组转染WWOX过表达质粒,NEG组转染空载质粒,空白对照组不做处理。采用细胞划痕实验测算各组细胞迁移率,以细胞迁移率表示细胞迁移能力。采用Transwell实验测算各组穿膜细胞数,以穿膜细胞数表示细胞侵袭能力。采用实时荧光定量PCR法检测各组细胞中WWOX、P73、E-cadherin、Vimentin mRNA,采用Western blotting法检测各组细胞中WWOX、P73、Ecadherin、Vimentin蛋白。结果实验组、NEG组、空白对照组细胞迁移率分别为50.42%±3.47%、74.81%±1.04%、77.82%±2.71%,其中实验组细胞迁移率与NEG组、空白对照组相比,P均<0.05。实验组、NEG组、空白对照组穿膜细胞数分别为(77.33±8.65)、(173.33±6.18)、(187.33±6.94)个,其中实验组穿膜细胞数与NEG组、空白对照组相比,P均<0.05。实验组WWOX、P73、E-cadherin mRNA和蛋白相对表达量均高于NEG组和空白对照组(P均<0.05),实验组Vimentin mRNA和蛋白相对表达量均低于NEG组和空白对照组(P均<0.05)。结论转染过表达WWOX质粒的GBC-SD细胞株迁移、侵袭能力降低,其机制可能与P73表达上调进而抑制GBC-SD细胞的上皮-间充质转化过程有关。
Objective To observe the changes of migration and invasion of gallbladder cancer cell line GBC-SD transfected with oxidoreductase(WWOX) plasmid overexpressing WW domain,and to explore their mechanism.Methods Gallbladder cancer cell line GBC-SD was divided into the experimental group,NEG group,and blank control group.The GBC-SD cell line in the experimental group was transfected with WWOX overexpression plasmid,the NEG group with empty plasmid,but the GBC-SD cell line in the blank control group was not treated.The cell migration rate was measured by cell scratch test,and the cell migration ability was expressed by cell migration rate.Trans well experiment was used to calculate the number of transmembrane cells in each group,and the number of transmembrane cells showed the ability of cell invasion.The mRNA levels of WWOX,p73,E-cadherin,and Vimentin were detected by real-time fluorescence quantitative PCR,and the protein levels of WWOX,p73,E-cadherin,and Vimentin were detected by Western blotting.Results The cell migration rates of the experimental group,NEG group,and blank control group were50.42%±3.47%,74.81%±1.04%,and 77.82%±2.71%,respectively.Significant differences were found in the cell migration rates between the experimental group and the NEG group,blank control group(all P<0.05).The number of transmembrane cells in the experimental group,NEG group and blank control group were 77.33±8.65,173.33±6.18,and 187.33±6.94,respectively-Significant difference was found in the number of transmembrane cells between the NEG group,blank control group and the experimental group(all P<0.05).The relative expression levels of WWOX,p73,and E-cadherin mRNA and protein in the experimental group were higher than those in the NEG group and the blank control group(all P<0.05),and the relative expression levels of Vimentin mRNA and protein in the experimental group were lower than those in the NEG group and the blank control group(all P<0.05).Conclusion The migration and invasion abilities of GBC-SD cell line transfected with overexpression of WWOX plasmid decreased,and the mechanism may be related to the up-regulation of p73 expression and it consequently inhibited the epithelial-mesenchymal transformation of GBC-SD cells.
作者
房立锐
魏东
雷学芬
佟鑫
徐冬杏
张文
FANG Lirui;WEI Dong;LEI Xuefen;TONG Xin;XU Dongxing;ZHANG Wen(Department of Hepatobiliary and Pancreatic Surgery,The Second Affiliated Hospital of Kunming Medical University,Kunming 650000,China;不详)
出处
《山东医药》
CAS
2022年第9期22-26,共5页
Shandong Medical Journal
基金
云南省科技厅—昆明医科大学应用基础研究联合专项资金面上项目(2019FE001-071)
云南省科技计划项目(2017FE468-057)
昆明医科大学研究生创新基金(2021S055)。
关键词
胆囊癌
GBC-SD细胞株
质粒
细胞迁移
细胞侵袭
上皮-间充质转化
gallbladder carcinoma
GBC-SD cell line
plasmid
cell migration
cell invasion
epithelial-mesenchymal transformation
