miR-203抑制食管鳞癌细胞迁移、侵袭及其分子机制探讨

miR-203 inhibits migration and invasion of esophageal squamous cancer cells and its molecular mechanism

目的:探究miR-203对食管鳞癌细胞(TR146、EC109)迁移、侵袭能力的影响及其分子机制。方法:检测miR-203在食管鳞癌细胞系中的表达水平,并通过转染miR-203激动剂agomir使TR146、EC109细胞稳定高表达miR-203,miR-203 agomir阴性对照组(NC)和无处理组(Blank)作为对照。通过划痕实验、Transwell实验检测miR-203对TR146、EC109细胞迁移、侵袭能力的影响。利用生物信息学分析miR-203潜在的靶基因,并通过双荧光素酶报告基因实验、实时定量PCR(qPCR)实验、Western blot实验验证miR-203靶基因。通过拯救实验探究miR-203是否通过抑制靶基因发挥作用。结果:与正常食管上皮细胞相比,miR-203在食管鳞癌细胞系中表达下调。在TR146、EC109细胞内将miR-203表达水平上调数倍,划痕实验证实miR-203能够抑制TR146、EC109细胞迁移能力,Transwell实验证实miR-203能够抑制TR146、EC109细胞侵袭能力。生物信息学、qPCR实验和Western blot实验表明LASP1(LIM and SH3 domain protein 1)是miR-203潜在的靶基因。拯救实验表明miR-203通过靶向抑制LASP1发挥抑制食管鳞癌细胞迁移、侵袭的作用。结论:miR-203能够抑制食管鳞癌细胞迁移、侵袭,并且该抑制作用可能通过miR-203靶向抑制LASP1介导,为食管鳞癌临床诊断和靶向治疗提供了理论依据。

Objective:To detect the effects of miR-203 on the migration and invasion capacity of TR146 and EC109 cells and explore its molecular mechanism.Methods:miR-203 agomir was transfected to upregulate miR-203 expression level,agomir NC group and Blank group were regarded as control group.qPCR was performed to examine the expression level of miR-203 in TR146 and EC109 cells,wound healing assays and Transwell chamber assays were used to study the effects of miR-203 on the migration and invasion of TR146 and EC109 cells.Then LASP1(LIM and SH3 domain protein 1)was predicted to be a potential target gene with bioinformatics and confirmed by means of dual luciferase reporter gene assays,qPCR and Western blot.Results:miR-203 agomir was efficiently expressed in TR146 and EC109 cells,wound healing assays and Transwell chamber assays results showed that upregulation of miR-203 can inhibit the migration and invasion of TR146 and EC109 cells.Dual luciferase reporter gene assays,qPCR and Western blot confirmed LASP1 to be a target gene of miR-203.These results obviously suggested the molecular mechanism by which miR-203 inhibits migration and invasion of TR146 and EC109 cells.Conclusion:miR-203 could inhibit migration and invasion of TR146 and EC109 cells,which may be explained by LASP1 being a target gene of miR-203.