miR-1246通过靶向GSK3β促进食管癌转移的机制  

miR-1246 promotes esophageal cancer metastasis by targeting GSK3β

作　　者：王明[1] 夏彦民 王文辰[1] 姜涛[1] 

机构地区：[1]第四军医大学唐都医院胸外科,陕西西安710038

出　　处：《现代肿瘤医学》2017年第9期1369-1374,共6页Journal of Modern Oncology

基　　金：陕西省自然科学基金A类(编号:2014A3)

摘　　要：目的:探讨miRNA-1246(miR-1246)促进食管癌细胞转移的作用及其机制。方法:Realtime-PCR法检测miR-1246在癌旁组织、食管癌组织、正常食管上皮和食管癌细胞系中的表达差异。Transwell侵袭实验观察转染miR-1246 mimics或inhibitors对食管癌细胞EC109转移能力的影响;裸鼠尾静脉转移实验观察稳定表达miR-1246对食管癌转移能力的影响;生物信息学分析miR-1246的候选靶基因为GSK3β,荧光素酶报告基因实验检测过表达miR-1246后EC109细胞中野生型和突变型GSK3β的荧光素酶活性。Western blot检测miR-1246对GSK3β蛋白表达的影响。结果:食管癌组织中的miR-1246表达显著高于癌旁组织(P<0.05);多个食管癌细胞中miR-1246的表达比正常食管上皮细胞Het-1A明显增高(P<0.05)。与阴性对照相比,miR-1246 mimics显著促进EC109细胞的侵袭能力(P<0.05)。反之,miR-1246 inhibitors明显抑制EC109细胞的侵袭能力(P<0.05);体内实验发现稳定过表达miR-1246明显升高EC109细胞的肺转移能力。荧光素酶报告基因结果证实miR-1246能够抑制GSK3β的3'-UTR区荧光素酶活性;转染miR-1246后,EC109细胞中的GSK3β蛋白水平明显低于对照组;miR-1246过表达显著抑制GSK3β-wt,而不能抑制GSK3β-mut的蛋白表达。结论:miR-1246能够通过靶向GSK3β促进食管癌转移,抑制miR-1246的表达或增加GSK3β的表达可能是抑制食管癌转移的有效手段。Objective：To investigate the role and mechanisms of miRNA-1246（miR-1246） in promoting esophageal cancer metastasis.Methods：Realtime-PCR was used to evaluate miR-1246 expression between esophageal cancer tissues and paired adjacent normal tissues,esophageal cancer cell lines and normal esophageal epithelial cells.After transfection of miR-1246 mimics or inhibitors,transwell invasion assay was applied to the effect of miR-1246 in esophageal cancer cell metastasis.Metastasis model in nude mice was used to observe the effect of miR-1246 overexpression in metastasis of esophageal cancer in vivo.Bioinformatics analysis revealed GSK3β as a candidate target genes of miR-1246.Luciferase activity of the luciferase reporter assay was used to detect the effect of miR-1246 on wild-type and mutant GSK3β in EC109 cells.Western blot was used to evaluate whether miR-1246 could affect protein expression of GSK3β and its mutant.Results：The expression of miR-1246 in esophageal cancer tissues was significantly elevated,compared with paired adjacent normal tissues（P〈0.05）.miR-1246 expression level in the esophageal cancer cells was significantly elevated,compared with normal esophageal epithelial cell Het-1A（P〈0.05）.miR-1246 mimics significantly increased invasion of EC109 cells,compared with the negative control（P〈0.05）.While miR-1246 inhibitors significantly inhibited invasion of EC109 cells（P〈0.05）.Stable expression of miR-1246 in EC109 cells demonstrated an increased invasion capacity in vivo.Luciferase reporter assay showed that miR-1246 inhibited luciferase activity of GSK3β by targeting its 3＇-UTR region.Western blot assay showed EC109 cells transfected with miR-1246 significantly decreased GSK3β expression,compared with the negative control.Western blotting showed that overexpression of miR-1246 significantly inhibited the expression of GSK3β-wt,but not GSK3β-mut.Conclusion：MiR-1246 promotes esophageal cancer metastasis by targeting GSK3β.Inhibition of miR-1246 expression or GSK3β

关 键 词：食管癌 转移 微小RNA miR-1246 GSK3Β 

分 类 号：R735.1[医药卫生—肿瘤]