miR-337靶向调节p53表达对结肠癌细胞自噬和迁移能力的影响  

Effects of miR-337 targeting p53 expression on autophagy and migration ability of colon cancer cells

作　　者：李磊[1] 张秘[2] 李文涛[3] 张代娟[4] 刘江月[4] LI Lei;ZHANG Mi;LI Wen-tao;ZHANG Dai-juan;LIU Jiang-yue(epartment of Anatomy,Weifang Medical College,Weifang 261053,China;Affiliated Hospital,Weifang Medical College,Weifang 261053,China;Functional Laboratories,Weifang Medical College,Weifang 261053,China;Departmentof Pathophysiology,Weifang Medical College,Weifang 261053,China)

机构地区：[1]潍坊医学院解剖学教研室,山东潍坊26105 [2]潍坊医学院附属医院,山东潍坊26105 [3]潍坊医学院机能学实验室,山东潍坊26105 [4]潍坊医学院病理生理教研室,山东潍坊26105

出　　处：《中国病理生理杂志》2019年第10期1791-1797,共7页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.8130068);山东省自然科学基金资助项目(No.ZR2015HL013)

摘　　要：目的:研究微小RNA-337(miR-337)对结肠癌细胞自噬及迁移能力的影响并从靶向调节p53表达的角度探讨其可能机制。方法:采用免疫组化方法检测结肠癌组织中beclin-1、LC3B和p53蛋白的表达,分析beclin-1和LC3B蛋白的表达与临床病理特征的相关性及p53与beclin-1和LC3B蛋白表达的相关性。小干扰RNA敲减p53基因表达后电镜观察结肠癌细胞株HCT116中自噬小体的形成,Western blot检测beclin-1和LC3B蛋白的表达。生物信息学预测筛选靶向p53的miRNAs并用RT-qPCR方法验证在HCT116细胞中的表达,萤光素酶报告检测miR-337对p53基因的调控作用。过表达miR-337后用Western blot检测p53及beclin-1和LC3B的表达,Transwell实验检测HCT116细胞的迁移能力。结果:与癌旁黏膜组织相比,beclin-1和LC3B蛋白在结肠癌组织中表达降低,与结肠癌的发生发展以及侵袭、转移显著相关。p53在结肠癌组织中表达升高,与beclin-1和LC3B蛋白的表达显著负相关。敲减p53基因的表达使beclin-1和LC3B蛋白表达上调;过表达miR-337和敲减p53蛋白表达可使beclin-1和LC3B的蛋白表达上调,HCT116细胞迁移能力降低(P<0.05)。结论:miR-337可促进结肠癌细胞自噬,抑制其迁移能力,其机制可能与靶向抑制p53表达有关。AIM:To investigate the effect of microRNA-337(miR-337)on the autophagy and migration ability of colon cancer cells,and to explore its possible mechanism involving targeting p53 expression.METHODS:The me-thod of immunohistochemistry was used to detect the protein expression of beclin-1,LC3B and p53 in colon cancer tissues.The correlations between the protein expression of beclin-1/LC3B and clinicopathological features,and the correlations between the protein expression of p53 and beclin-1/LC3B were analyzed.After knock-down of p53 expression by small interfering RNA,the formation of autophagiosomes was observed under electron microscope in colon cancer cell line HCT116,and the protein expression of beclin-1 and LC3B was determined by Western blot.The miRNAs targeting p53 were predicted and screened by bioinformatics,and their expression in HCT116 cells was verified by RT-qPCR.Luciferase reporter assay was used to detect the regulatory effect of miR-337 on p53 gene.The protein expression of p53,beclin-1 and LC3B was determined by Western blot,and the migration ability of HCT116 cells after miR-337 over-expression was detected by Transwell assay.RESULTS:The protein expression of beclin-1 and LC3B in the colon cancer tissues was decreased,which was significantly related to the occurrence,development,invasion and metastasis of colon cancer.The expression of p53 was increased in the colon cancer tissues,which was negatively correlated with the protein expression of beclin-1 and LC3B.Knock-down of p53 gene expression increased the protein expression of beclin-1 and LC3B(P<0.05).Over-expression of miR-337 down-regulated the expression of p53,up-regulated the protein expression of beclin-1 and LC3B,and decreased the migration ability of HCT116 cells(P<0.05).CONCLUSION:miR-337 promotes autophagy and inhibits migration ability of colon cancer cells,and the mechanism may be related to targeted inhibition of p53 expression.

关 键 词：结肠癌 微小RNA-337 P53蛋白 自噬 细胞迁移 

分 类 号：R730.23[医药卫生—肿瘤] R735.3[医药卫生—临床医学]