MiR-412-3p表达水平与鼻咽癌发病的关系及其对鼻咽癌细胞株CNE1顺铂敏感性的影响  

Relationship between the expression level of mi R-412-3p and the incidence of nasopharyngeal carcinoma and its effect of cisplatin sensitivity on nasopharyngeal carcinoma cell line CNE1

作　　者：刘惺 喻泊遥 王慷慨 马恒 梁琳 周艳宏 LIU Xing;YU Boyao;WANG Kangkai;MAHeng;LIANG Lin;ZHOU Yanhong(Functional Experiment Center,School of Basic Medicine,Central South University,Changsha 410078;Department of Pathophysiology,School of Basic Medicine,Central South University,Changsha 410078;Cancer Research Institute,School of Basic Medicine,Central South University,Changsha 410078,China)

机构地区：[1]中南大学基础医学院机能学实验中心,长沙410078 [2]中南大学基础医学院病理生理学系,长沙410078 [3]中南大学基础医学院肿瘤研究所,长沙410078

出　　处：《中南大学学报（医学版）》2020年第7期745-751,共7页Journal of Central South University ：Medical Science

基　　金：湖南省自然科学基金重点项目(12JJ2044)。

摘　　要：目的:探讨miR-412-3p及乳运铁蛋白(lactotransferrin,LTF)在鼻咽癌组织及癌旁组织中的表达差异并观察干预miR-412-3p能否改善鼻咽癌细胞株CNE1的顺铂敏感性。方法:分别采用实时反转录聚合酶链反应(RT-qPCR)及蛋白质印迹法检测收集的20例配对的鼻咽癌组织及癌旁组织中miR-412-3p及LTF的表达水平;MTT法及克隆形成实验检测miR-412-3p抑制剂、顺铂处理对CNE1细胞活性及克隆形成能力的影响;蛋白质印迹法检测不同处理对CNE1细胞ERK1/2,STAT3,AKT信号通路的影响。结果:与配对的癌旁组织相比,鼻咽癌组织中miR-412-3p的表达明显升高,伴随LTF表达的显著降低(均P<0.01);在体外培养的CNE1细胞中,不同浓度的miR-412-3p抑制剂可显著抑制miR-412-3p及磷酸化AKT蛋白的表达,同时促进LTF蛋白的表达(P<0.05);预先使用200 nmol/L miR-412-3p抑制剂处理CNE1细胞24 h后,可明显增加20μg/mL顺铂对CNE1细胞活性及克隆形成能力的抑制(P<0.01)。结论:鼻咽癌组织中miR-412-3p的高表达及LTF的低表达可能与鼻咽癌的发病有关;利用miR-412-3p抑制剂降低CNE1细胞中miR-412-3p的水平可增加CNE1细胞对顺铂的敏感性。Objective:To investigate the differences in expressions of miR-412-3p and lactotransferrin(LTF)in nasopharyngeal carcinoma tissues and adjacent tissues,and to observe whether the intervention of miR-412-3p can improve the cisplatin sensitivity of nasopharyngeal carcinoma cell line CNE1.Methods:The expression levels of miR-412-3p and LTF in 20 paired nasopharyngeal carcinoma tissues and adjacent tissues were detected by RT-q PCR and Western blotting.The effects of miR-412-3p inhibitors and cisplatin treatment on CNE1 activity and clonogenesis were detected by MTT and colony formation assay.The effects of different treatments on ERK1/2,STAT3 and AKT signaling pathways in CNE1 cells were detected by Western blotting.Results:Compared with paired paracancer tissues,the expression of miR-412-3p in nasopharyngeal carcinoma tissues was significantly increased,accompanied by a significant decrease in LTF expression both(P<0.01).In cultured CNE1 cells,miR-412-3p inhibitors of different concentrations can significantly inhibit the expression of miR-412-3p and p-AKT,and promote the expression of LTF protein(P<0.05).Pretreatment of CNE1 cells with 200 nmol/L miR-412-3p inhibitor for 24 h significantly enhanced the inhibition of 20μg/m L cisplatin on CNE1 cell activity and clonogenesis(P<0.01).Conclusion:The elevated expression of miR-412-3p and the decreased expression of LTF in nasopharyngeal carcinoma may be related to the incidence of nasopharyngeal carcinoma.Using miR-412-3p inhibitors to reduce the level of miR-412-3p in CNE1 cells can increase the sensitivity of CNE1 cells to cisplatin.

关 键 词：鼻咽癌 miR-412-3p 乳运铁蛋白 顺铂 蛋白激酶B 

分 类 号：R739.63[医药卫生—肿瘤]