摘要
目的探讨去泛素化酶3(ubiquitin-specific protease 3,USP3)在肺癌组织中的表达及对肺癌细胞上皮间质转化、迁移和侵袭的影响。方法利用反转录-多聚酶链式反应(reverse transcription-polymerase chain reaction,RT-PCR)和Western blot等方法检测87例肺癌患者肺癌组织和癌旁组织中USP3的表达情况。将人肺癌细胞(A549)分为3组:未做处理的对照组,转染control siRNA的siRNA-control组以及转染干扰USP3的siRNA-USP3组,RT-PCR和Western blot方法检测3组细胞中USP3水平,以观察干扰效果;利用Western blot方法检测上皮细胞-间质细胞转化(epithelial-mesenchymal transition,EMT)相关的E-钙黏蛋白(epithelia cadherin,E-cadherin)及波形蛋白(Vimentin)的表达情况;划痕愈合实验检测细胞迁移能力;Transwell小室法检测细胞侵袭能力。结果USP3在肺癌组织中的表达水平显著高于癌旁组织(P<0.05)。siRNA-USP3组的A549细胞中USP3的转录和表达水平显著低于对照组和siRNA-control组(P<0.05)。E-cadherin在siRNA-USP3组的水平显著高于对照组和siRNA-control组(P<0.05),Vimentin在siRNA-USP3组的水平显著低于对照组和siRNA-control组(P<0.05)。与其他2组相比,siRNA-USP3组的划痕愈合率以及侵袭能力显著降低(P<0.05)。结论USP3在肺癌患者体内呈高水平表达,可促进A549细胞的上皮间质转化、迁移和侵袭能力。
Objective To investigate the expression of ubiquitin-specific protease 3(USP3)in lung cancer tissues and its effects on lung cancer cell epithelial-mesenchymal transition(EMT),migration and invasion.Methods The expression levels of USP3 in lung cancer tissues and the corresponding normal tissues of 87 patients with lung cancer were detected by Reverse Transcription-Polymerase Chain Reaction(RT-PCR)and Western Blot.The human lung cancer cells(A549)were divided into three groups:the cells without any treatment were served as control group,and the cells were transfected with control siRNA were served as siRNA-control group,and the cells were transfected with siRNA targeting USP3 were served as siRNA-USP3 group.The expression levels of USP3 were detected RT-PCR and Western blot to observe their interference effects.And the expression levels of E-cadherin and Vimentin were detected by Western Blot to observe the EMT,and cell migration was tested by scratch healing experiment,moreover,cell invasion was tested by Transwell experiment.Results The expression levels of USP3 in lung cancer tissues were significantly higher than those in corresponding normal tissues(P<0.05).The transcription and expression levels of USP3 in siRNA-USP3 group were significantly lower than those in siRNA-control group and control group(P<0.05).The expression levels of E-cadherin in siRNA-USP3 group were significantly higher than those in siRNA-control group and control group(P<0.05),however,the expression levels of Vimentin siRNA-USP3 group were significantly lower than those in siRNA-control group and control group(P<0.05).In addition the scratch healing rate and cell invasion ability in siRNA-USP3 group were significantly lower than those in siRNA-control group and control group(P<0.05).Conclusion USP3 is highly expressed in patients with lung cancer,which can promote EMT,and migration and invasion ability of A549 cells.
作者
肖蓓
邓小婧
XIAO Bei;DENG Xiaojing(Meishan Tumor Hospital,Sichuan,Meishan 620010,China)
出处
《河北医药》
CAS
2020年第7期1012-1015,共4页
Hebei Medical Journal
关键词
去泛素化酶3
肺癌细胞
上皮间质转化
迁移
侵袭
ubiquitin-specific protease 3
lung cancer cells
epithelial-mesenchymal transition
migration
invasion
