miR-106b对鼻咽癌细胞凋亡的促进和对增殖的抑制作用  

miR-106b induces the apoptosis and inhibits the proliferation of nasopharyngeal carcinoma cells

作　　者：严达忠[1] 邓世山[2] 李四军[1] 严晨 甘卫刚[1] 

机构地区：[1]川北医学院附属医院耳鼻咽喉头颈外科,四川南充637000 [2]川北医学院人体解剖教研室,四川南充637000 [3]大连医科大学第一临床学院,辽宁大连116000

出　　处：《西安交通大学学报（医学版）》2017年第6期851-856,共6页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：四川省科技厅重点项目资助(No.14ZA0198)~~

摘　　要：目的探讨miR-106b对鼻咽癌细胞凋亡和增殖的影响。方法应用miRNA芯片分析获得鼻咽癌组织和癌旁正常组织表达差异的miRNA,TaqMan miRNA检测试剂盒和实时荧光定量PCR分别检测miR-106和RhoC mRNA在鼻咽癌和癌旁组织中的表达,用miRnada预测miR-106b和靶基因的结合位点,采用双荧光素酶验证靶基因,Western blot检测miR-106b调控靶基因RhoC的表达水平。用Annexin V-PI和TUNEL检测miR-106b对鼻咽癌细胞凋亡的影响,用MTT检测miR-106b对鼻咽癌细胞增殖的影响。结果 miRNA芯片分析发现鼻咽癌组织中miR-106b的表达较癌旁正常组织低。RT-PCR结果显示miR-106b在鼻咽癌组织中表达减少(P<0.05),RhoC在鼻咽癌组织中表达增加(P<0.05),miR-106b和RhoC在鼻咽癌组织中表达呈负相关(r=―0.586 6,P<0.001)。荧光素酶报告基因实验结果显示miR-106b组荧光素酶活性低于空质粒组(P<0.05),Western blot结果显示miR-106b组鼻咽癌细胞中RhoC的表达较空质粒组降低(P<0.05)。Annexin V-PI和TUNEL结果显示,miR-106b组鼻咽癌细胞凋亡较空质粒组增加(P<0.05);MTT结果显示miR-106b组鼻咽癌细胞增殖能力较空质粒组降低(P<0.05)。结论 miR-106b可能通过下调RhoC的表达诱导鼻咽癌细胞凋亡并抑制鼻咽癌细胞增殖。Objective To investigate the effect of miR-106 b on the apoptosis and proliferation of nasopharyngeal carcinoma（NPC） cells. Methods We analyzed differences in miRNA expression in nasopharyngeal carcinoma and adjacent normal tissues with miRNA microarray.TaqMan miRNA detection kit and Real-time fluorescence quantitative PCR were used to detect the expressions of miR-106 and RhoC mRNA in nasopharyngeal carcinoma and adjacent tissues.The miR-106 b and target gene binding sites were predicted with miRnada.The target gene was verified by double luciferase.Western blot was used to detect the expression of RhoC regulated by miR-106 b.Annexin and TUNEL were used to detect the effect of miR-106 b on the apoptosis of nasopharyngeal carcinoma cells;the effect of miR-106 b on the proliferation of nasopharyngeal carcinoma cells was detected by MTT assay.Results miRNA microarray analysis showed that the expression of miR-106 b was lower in NPC tissues than in adjacent normal tissues.The results of RT-PCR showed that the expression of miR-106 b in nasopharyngeal carcinoma was decreased（P〈0.05）while the expression of RhoC was increased in nasopharyngeal carcinoma（P〈0.05）.The expressions of miR-106 b and RhoC in NPC were negatively correlated（r=-0.586 6,P〈0.001）.The results of luciferase reporter assay showed that the activity of luciferase in miR-106 b group was lower than that in empty plasmid group（P〈0.05）.The results of Western blot showed that miR-106 b could decrease the expression of RhoC in NPC tissues（P〈0.05）.Annexin V-PI and TUNEL showed that the apoptosis of nasopharyngeal carcinoma cells was significantly higher in miR-106 group than in empty plasmid group（P〈0.05）.MTT results showed that the proliferation of nasopharyngeal carcinoma cells in miR-106 b group was lower than that in empty plasmid group（P〈0.05）.Conclusion miR-106 b may induce the apoptosis of nasopharyngeal carcinoma cells and inhibit the proliferation of nasopharyngeal carcinoma cells by down-regulating the e

关 键 词：miR-106b 鼻咽癌 凋亡 增殖 RHOC 

分 类 号：R543.3[医药卫生—心血管疾病]