摘要
目的探讨泛素特异性蛋白酶9X(USP9X)shRNA对非小细胞肺癌(NSCLC)A549细胞侵袭和凋亡影响.方法USP9X shRNA慢病毒感染A549细胞,Realtime PCR和Western blot检测下调效果,MTT检测细胞增殖,细胞克隆实验测定克隆能力,流式细胞术测定凋亡,Transwell小室测定侵袭和迁移,免疫印迹检测KI-67、PCNA、C-Caspase-3、C-Caspase-9、MMP-2、MMP-9、E-Cadherin、Vimentin蛋白表达.结果USP9X shRNA慢病毒感染后的A549细胞中USP9X表达水平下降,细胞增殖能力、克隆形成能力降低,细胞凋亡水平增加,细胞侵袭和迁移能力下降,细胞中KI-67、PCNA、MMP-2、MMP-9蛋白水平下降,C-Caspase-3、C-Caspase-9蛋白水平升高,E-Cadherin蛋白水平升高,Vimentin蛋白水平降低.结论USP9X shRNA抑制NSCLC A549细胞侵袭并诱导凋亡.
Objective To investigate the effect of ubiquitin-specific protease 9X(USP9X)shRNA on the invasion and apoptosis of non-small cell lung cancer cell line A549.Methods USP9X shRNA lentivirus was used to infect A549 cells.Realtime PCR and Western blot were used to determine the down-regulation effect.MTT assay was used to examine cell proliferation.Cell cloning assay was used to determine the cloning ability.Flow cytometry was used to determine the apoptosis.Transwell chamber assay was used to measure the invasion and migration.Western blot was used to determine the protein expression levels of KI-67,PCNA,C-Caspase-3,C-Caspase-9,MMP-2,MMP-9,E-Cadherin and Vimentin.Results After infected by the USP9X shRNA lentivirus,the expression level of USP9X in A549 cells decreased;the ability of cell proliferation and colony formation decreased;the apoptosis level increased,whereas the cell invasion and migration ability decreased;the protein levels of KI-67,PCNA,MMP-2 and MMP-9 decreased;the protein levels of C-Caspase-3 and C-Caspase-9 increased;the protein level of E-Cadherin increased;and the protein level of Vimentin decreased.Conclusion USP9X shRNA inhibits the invasion of NSCLC A549 cell line and thereby induces its apoptosis.
作者
李志杰
韦海涛
张伟华
李丽
Li Zhijie;Wei Haitao;Zhang Weihua;Li Li(Second Ward,Department of Respiratory and Critical Care Medicine,Shangqiu First Municipal People’s Hospital,Shangqiu,Henan 476100,China;Department of Thoracic Surgery,Huaihe Hospital of Henan University,Kaifeng,Henan 475004,China;Department of Internal Medicine,School of Nursing and Health,Henan University,Kaifeng,Henan 475004,China)
出处
《中华生物医学工程杂志》
CAS
2019年第4期460-465,共6页
Chinese Journal of Biomedical Engineering
基金
河南省教育厅高等学校重点科研项目(18B310014)。
