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

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=？D0.5866，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。
ABSTRACT: Objective To investigate the effect of miR-106b 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-106b 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-106b. Annexin and TUNEL were used to detect the effect of miR-106b on the apoptosis of nasopharyngeal carcinoma cells; the effect of miR-106b on the proliferation of nasopharyngeal carcinoma cells was detected by MTT assay. Results miRNA microarray analysis showed that the expression of miR-106b was lower in NPC tissues than in adjacent normal tissues. The results of RT-PCR showed that the expression of miR-106b 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-106b and RhoC in NPC were negatively correlated (r=-0.5866, P<0.001). The results of luciferase reporter assay showed that the activity of luciferase in miR-106b group was lower than that in empty plasmid group (P<0.05). The results of Western blot showed that miR-106b 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-106b group was lower than that in empty plasmid group (P<0.05). Conclusion miR-106b may