环状RNAcirc-ccnb1通过抑制细胞周期蛋白核转位对食管鳞癌增殖和迁移的影响  

Circular RNA affect the proliferation and migration of esophageal squamous cell carcinoma by inhibiting cell cycle protein nuclear translocation

作　　者：杨瑜 刘耀河 秦宁 谭柳 宋锐[1] 侯歌[1] 袁金金[1] 张艳 刘宗文[1] YANG Yu;LIU Yaohe;QIN Ning;TAN Liu;SONG Rui;HOU Ge;YUAN Jinjin;ZHANG Yan;LIU Zongwen(Department of Radiation Therapy, The Second Affiliated Hospital of Zhengzhou University, Zhengzhou 450014, China;Department of Pharmacology, School of Basic Medical Sciences, Medical College of Zhengzhou University, Zhengzhou University, Zhengzhou 450014, China)

机构地区：[1]郑州大学第二附属医院放射治疗科,河南郑州450014 [2]郑州大学基础医学院药理学教研室,河南郑州450014

出　　处：《西部医学》2021年第8期1101-1105,共5页Medical Journal of West China

基　　金：国家重点研发计划(2016YFC0105713);省部科技共建项目(201401009)。

摘　　要：目的探讨环状RNA circ-ccnb1对细胞周期蛋白(cyclinb1、ccnb1)的作用,以及对食管鳞癌增殖和迁移的影响。方法①收集60例食管鳞癌组织及癌旁正常组织标本,按癌组织及正常组织分为两组,通过免疫组化检测两组组织中ccnb1的表达,并分析食管鳞癌中ccnb1表达与临床病理因素(淋巴结转移、病理分期)之间的相关性。②培养食管鳞癌细胞,并分为两组:正常食管上皮细胞(Het1-A)和食管鳞癌细胞109(EC109)。采用蛋白免疫印迹法(WB)检测两组中ccnb1蛋白的表达。③用携带circ-ccnb1的质粒转染EC109,检测其中ccnb1的核转位情况,将细胞分为3组:未转染组(Blank组)、转染空载体组(Vector组)及转染circ-ccnb1组(cir-ccnb1组)。转染成功后进行核桨分离,采用WB检测转移至胞核内的ccnb1蛋白量的变化,细胞增殖实验(CCK-8法)及划痕实验检测细胞增殖率、迁移率。结果食管鳞癌组织中ccnb1蛋白的表达高于正常组织,EC109细胞中ccnb1蛋白的表达高于Het1-A细胞(均P<0.05)。与Blank组相比,cir-ccnb1组细胞中ccnb1蛋白的核转位水平明显降低,细胞增值率、迁移率降低(均P<0.05)。结论cir-ccnb1可以通过抑制ccnb1在食管鳞癌细胞中的核转位进而影响食管鳞癌的发展,即抑制癌细胞增殖和迁移。Objective To investigate the relationship between circular RNA circ-ccnb1 and cyclinb1 and its effect on the development(proliferation and migration)of esophageal cancer.Methods 60 cases of esophageal squamous cell carcinoma and adjacent normal tissues were collected and divided into two groups.The expression of ccnb1 in the two groups was detected by immunohistochemistry,and the correlation between the expression of ccnb1 and clinicopathological factors(lymph node metastasis,pathological stage)was analyzed.Esophageal squamous cell carcinoma cells were cultured and divided into two groups:normal esophageal epithelial cells(Het1-A)and esophageal squamous cell carcinoma cells 109(Ec109).The expression of ccnb1 protein was detected by Western blot.The cells were divided into three groups:blank group,vector group and cir ccnb1 group.WB was used to detect the change of ccnb 1 protein transferred into the nucleus.Cell proliferation rate(CCK-8 method)and cell migration rate were detected by cell proliferation assay(CCK-8 method)and scratch test.Results The expression level of ccnb1 protein in esophageal squamous cell carcinoma tissues was higher than that in normal tissues(P<0.05),and the expression level of ccnb1 protein in EC109 cells was higher than that in Het1-A cells(P<0.05).Compared with the control group,the nuclear translocation level of ccnb1 protein in circ-ccnb1 cells was significantly lower(P<0.001),and the cell proliferation rate and migration rate were both lower(P<0.05).Conclusion Circ-ccnb1 can affect the development of esophageal squamous cell carcinoma by inhibiting the nuclear translocation of ccnb1 in esophageal squamous cell carcinoma cells,that is,inhibiting proliferation and migration.

关 键 词：circ-ccnb1 ccnb1 食管鳞癌 增殖 迁移 

分 类 号：R735.1[医药卫生—肿瘤]