微RNA-429通过抑制Bmi-1表达提高食管鳞癌细胞的化疗敏感性  

Micro RNA-429 promotes chemotherapeutic sensitivity of esophageal squamous cell carcinoma cells by inhibiting Bmi-1 expression

作　　者：马鸣[1] 栾亦然 白函瑜 赵连梅[2] 刘丽华[3] 单保恩[2] MA Ming;LUAN Yiran;BAI Hanyu;ZHAO Lianmei;LIU Lihua;SHAN Baoen(Department of Laboratory,Fourth Affiliated Hospital of Hebei Medical University,Shijiazhuang 050011,Hebei Province,China;Research Center,Fourth Affiliated Hospital of Hebei Medical University,Shijiazhuang 050011,Hebei Province,China;Department of Tumor Immunity,Fourth Affiliated Hospital of Hebei Medical University,Shijiazhuang 050011,Hebei Province,China)

机构地区：[1]河北医科大学第四医院检验科,河北石家庄050011 [2]河北医科大学第四医院科研中心,河北石家庄050011 [3]河北医科大学第四医院肿癯免疫科,河北石家庄050011

出　　处：《肿瘤》2019年第12期993-1003,共11页Tumor

基　　金：国家自然科学基金资助项目（编号：81703073);河北省自然科学基金资助项目（编号：H2018206115)~~

摘　　要：目的:研究微RNA(microRNA,miR)-429在食管鳞状细胞癌(esophageal squamous cell carcinoma,ESCC)中表达及其与患者临床特征和预后之间的关系,并且探讨miR-429对ESCC细胞化疗敏感性的调控作用及其可能的机制。方法:选取2010年1月-12月于河北医科大学第四医院就诊的60例ESCC患者的癌组织及癌旁组织标本,采用实时荧光定量PCR法检测ESCC组织中miR-429的表达水平,并分析其与患者临床特征及预后的关系。将miR-429模拟物或抑制物分别转染ESCC细胞株Eca109和TE-13后,MTT法检测miR-429过表达或沉默对细胞增殖活性及顺铂(cisplatin,CDDP)敏感性的影响,同时FCM法检测细胞凋亡水平的变化。双荧光素酶报告基因系统分析miR-429是否直接调控B细胞特异性莫洛尼鼠白血病病毒插入位点1(B-cell specific moloney murine leukemia virus insertion site-1,Bmi-1)基因表达,蛋白质印迹法检测细胞中Bmi-1及P-糖蛋白(P-glycoprotein,P-gp)表达水平的变化。结果:ESCC组织中miR-429表达水平较癌旁组织明显降低(P <0.01)。而且miR-429低表达的ESCC患者往往肿瘤体积较大,临床分期较晚,更易发生淋巴结转移,同时患者生存期较短(P值均<0.05)。过表达miR-429的Eca109和TE-13细胞增殖活性明显降低,同时对CDDP的敏感性明显增强,细胞凋亡率明显升高,表达Bmi-1及P-gp的水平明显降低(P值均<0.05);敲低miR-429表达的Eca109和TE-13细胞增殖活性明显升高,同时对CDDP的敏感性明显减弱,细胞凋亡率明显降低,表达Bmi-1及P-gp的水平明显升高(P值均<0.05)。双荧光素酶报告基因系统分析显示,Bmi-1是miR-429的靶基因,二者表达水平之间存在负相关性(r=-0.340 4,P <0.05)。结论:ESCC组织中miR-429低表达与患者肿瘤进展及预后较差密切相关。同时,miR-429可能具有增强ESCC细胞对CDDP化疗敏感性的作用,且其作用机制可能与调控Bmi-1表达有关。Objective: To study the expression of microRNA-429(miR-429) in esophageal squamous cell carcinoma(ESCC) tissues and its relationship with the clinical characteristics and prognosis of patients, as well as to explore the regulatory effect of miR-429 on the chemotherapeutic sensitivity of ESCC cells and the underlying mechanism.Methods: The samples of tumor tissues and paracancerous tissues were collected from 60 cases of ESCC patients from January 2010 to December 2010. The expression level of miR-429 in tumor tissues and paracancerous tissues was detected by real-time fluorescent quantitative PCR, as well as its relationship with the clinical feathers of ESCC patients was analyzed. After the miR-429 mimic or miR-429 inhibitor was transfetcted into Eca109 and TE-13 cells, the proliferation and chemotherapy sensitivity of ESCC cells were measured by MTT assay, the apoptosis of ESCC cells treated with cisplatin(CDDP) was detected by FCM. A dual-luciferase reporter assay was performed to detect whether B-cell specific moloney murine leukemia virus insertion site-1(Bmi-1) was a target gene of miR-429. The expressions of Bmi-1 and P-glycoprotein(P-gp) proteins in ESCC cells were analyzed by Western blotting.Results: The expression level of miR-429 in ESCC tissues was significantly higher than that in the paracancerous tissues(P < 0.01). For the ESCC tissues with miR-429 low expression, the size of tumor was bigger, the clinical stage of tumor was later, the lymph node metastasis was more likely to occure, and the survival time of ESCC patients was shorter as compared with the miR-429 higher expression group. After overexpressing miR-429, the proliferation viability of Eca109 and TE-13 cells were suppressed obviously, the sensitivity to CDDP was enhanced, the apoptosis rate was increased, while the expression levels of Bmi-1 and P-gp were significantly down-regulated(all P < 0.05). After blocking miR-429 expression, the proliferation viability of Eca109 and TE-13 cells were promoted obviously, the sensitivity to CDDP wa

关 键 词：食管肿瘤 微RNAS 顺铂 预后 miR-429 

分 类 号：R735.1[医药卫生—肿瘤]