摘要
目的探讨死亡相关蛋白激酶1(DAPK)在人食管鳞癌组织及EC9706细胞中的表达及其对食管鳞癌转移侵袭的作用。方法采用免疫组织化学法检测人食管鳞癌组织及癌旁正常组织中DAPK的表达。培养人食管癌EC9706细胞,分别用pReceiver-M29-DAPK质粒和pReceiver-M29转染细胞,以未转染的细胞作为对照组,各组细胞培养48h。应用蛋白印迹法(Western blot)检测各组细胞中DAPK蛋白表达的变化,划痕修复实验检测细胞迁移能力,Transwell小室侵袭实验检测细胞侵袭能力。结果人食管鳞癌组织DAPK的阳性率及DAPK的阳性区域面积明显高于癌旁正常组织(P<0.05)。在未转染的EC9706细胞中和空载脂质体转染的EC9706细胞中未见明显DAPK蛋白表达,DAPK蛋白在转染后的EC9706细胞中高表达(P<0.05)。与未转染的EC9706细胞比较,pReceiverM29-DAPK转染的EC9706细胞迁移率及细胞侵袭数目明显升高(P<0.01),未转染的EC9706细胞与空载脂质体转染的EC9706细胞迁移率及细胞侵袭数目差异无统计学意义(P>0.05)。结论 DAPK在人食管鳞癌组织中高表达,DAPK高表达后促进了EC9706细胞转移和侵袭。
Objective To investigate the expression of death-associated protein kinase(DAPK)in human esophageal squamous carcinoma tissue and EC9706 cells and its role on the metastasis and invasion of esophageal squamous carcinoma.Methods The immunohistochemistry was used to detect the expression of DAPK in human esophageal squamous carcinoma tissue and paracancerous normal tissues.Human esophageal carcinoma EC9706 cells were cultured and transfected with pReceiver-M29-DAPK plasmid and negative control(pReceiver-M29)respectively.The untransfected cells were used as the control group,and the cells in each group were cultured for 48 h.Western blot was used to detect the changes of DAPK protein expression in the cells of each group.The scratch repair test was used to detect the migration ability of EC9706 cells.The Transwell chamber invasion was used to detect the invasion ability of EC9706 cells.Results The positive rate of DAPK and the area of DAPK positive region in human esophageal squamous carcinoma tissue were significantly higher than those in paracancerous normal tissues(P<0.05).In untransfected EC9706 cells and in EC9706 cells transfected with empty liposomes,no significant DAPK expression was observed,and DAPK protein showed significantly high expression in EC9706 cells after transfection(P<0.05).Compared with nontransfected EC9706 cells,EC9706 cells transfected with pReceiverM29-DAPK significantly increased the migration rate of EC9706 cells and the number of cell invasion(P<0.01),and the cellular migration rate and cellular invasive number had no statistically significant difference between non-transfected EC9706 cells and EC9706 cells transfected with empty liposomes(P>0.05).Conclusion DAPK is highly expressed in human esophageal squamous carcinoma,and the high expression of DAPK promotes the migration and invasion of EC9706 cells.
作者
贾敬周
孙继伟
袁五营
侯智亮
王文波
赵正国
JIA Jingzhou;SUN Jiwei;YUAN Wuying;HOU Zhiliang;WANG Wenbo;ZHAO Zhengguo(Department of Minimally Invasive Surgery,Henan Provincial Thoracic Hospital,Zhengzhou,Henan 450000,China;Department of General Surgery,Zhengzhou Municipal Seventh People's Hospital,Zhengzhou Henan 450002,China)
出处
《重庆医学》
CAS
2019年第6期925-928,共4页
Chongqing medicine
基金
河南省科技发展计划项目(162102310219)
