miR-196b作为一种喉鳞癌预后因子通过靶向基因对于肿瘤细胞凋亡、增殖作用的研究  

miR-196b affects the progression and prognosis of human LSCC through targeting WDR37

作　　者：姜丽[1] 李玉茹[1] Jiang Li;Li Yuru(Department of Otolaryngology Head and Neck Surgery,the First Affiliated Hospital of Harbin Medical University,Harbin 150001,China)

机构地区：[1]哈尔滨医科大学附属第一医院耳鼻喉科,150001

出　　处：《国际遗传学杂志》2020年第2期76-81,共6页International Journal of Genetics

摘　　要：目的探讨miRNA-196b以及WDR37对于人喉鳞状细胞癌(laryngeal squamous cell carcinoma,LSCC)的作用及其潜在的作用机制。方法通过肿瘤基因图谱分析miR-196b和WDR37在LSCC患者中的表达以及两者的相关性。在Tu868细胞低表达miR-196b或过表达WDR37通过CCK-8、AO/EB以及克隆形成检测细胞活性,凋亡以及增殖的作用,通过荧光素酶基因报告研究miR-196b与WDR37的靶向关系,Western blot方法检测转染miR-196b后增殖相关蛋白的变化。结果miR-196b在LSCC中的表达量明显高于正常组(P=0.028),且高表达miR-196b的患者生存率明显低于低表达组(P=3.000×10^-4),WDR37在LSCC中的表达量明显低于正常组(P=0.048)。低表达miR-196b的LSCC患者生存时间明显高于高表达(P=0.001);miR-196b与WDR37在LSCC患者中表达呈负相关(P=5.930×10^-12)。miR-196b inhibitor可使WDR37的表达增加(P=0.032、0.042),miR-196b mimic可使WDR37的表达降低(P=0.029、0.043)。WDR37是miR-196b的直接靶点。miR-196b inhibitor能够明显抑制细胞活性(P=0.042);而miR-196b mimic能够使细胞活性明显增加(P=0.031)。结果显示miR-196b inhibitor能够诱导Tu868细胞凋亡,同时Ki67实验的结果也说明,miR-196b inhibitor能够明显的抑制Tu868细胞增殖。结论miR-196b过表达抑制人LSCC中的WDR37,导致肿瘤发展预后不良。miR-196b是一种潜在的预后评估标志物。Objective To explore the effect of miR-196b and WDR37 on the biological features of human laryngeal squamous cell carcinoma(LSCC)and its potential mechanism.Methods miR-196b and WDR37 expressions were determined in TCGA,and the targeting relation between miR-196b and WDR37 was verified by dual-luciferase reporter system.In vitro,Tu868 cells were divided into the Control,miR-196b inhibitors,miR-NC,WDR37 and miR-196b mimics+WDR37 groups.The miR-196b and WDR37 expressions were determined by qRT-PCR or/and Western blot,and their biological features were determined by CCK-8,AO/EB and colony assays.Results miR-196b in LSCC was significantly higher than that in normal group(P=0.028),and the survival rate of patients with high expression of miR-196b was significantly lower than that in low expression group(P=3.000×10^-4),and the expression of WDR37 in LSCC was significantly lower than that in normal group(P=0.048).The survival time of LSCC patients with low expression of miR-196b was significantly higher than that with high expression(P=0.001);the expression of miR-196b and wdr37 was negatively correlated in LSCC patients(P=5.930×10^-12).MiR-196b inhibitor increased the expression of WDR37(P=0.032,0.042),and miR-196b mimic reduced the expression of WDR37(P=0.029,0.043).WDR37 is the direct target gene of miR-196b.MiR-196b inhibitor significantly inhibited cell growth(P=0.042),while miR-196b mimic significantly increased cell growth(P=0.031).The results showed that miR-196b inhibitor can induce apoptosis of TU868 cells,and Ki67 staining also showed that miR-196b inhibitor can significantly inhibit the proliferation of Tu868 cells.Conclusion LSCC patients were characterized by up-regulation of miR-196b and down-regulation of WDR37,which are correlated with the major clinical features and prognosis.Inhibiton of miR-196b may suppress proliferation and promote apoptosis of Tu868 cells via targeting WDR37.

关 键 词：miR-196b 预后 活性 人喉鳞状细胞癌 

分 类 号：R73[医药卫生—肿瘤]