目的
微RNA（MicroRNAs，miRNAs）是一组与肺癌等不同癌症相关的非编码小RNA。本文旨在探讨循环mirna的表达谱及其在肺癌发生发展中的作用。

材料和方法
采用miRNAs法检测肺癌患者循环miRNA水平。然后通过搜索基因数据库，预测异常表达mirna的靶基因。以miR-1246和miR-1246抑制剂转染A549细胞为基础，进一步测定miR-1246在肺癌细胞体外的上皮间质转移（EMT）、迁移和侵袭能力中的作用。最后，分别用免疫荧光法和westernblot法检测miR-1246对糖原合成酶激酶-3β（GSK-3β）/β-连环素途径的影响。

结果
我们发现肺癌患者血清中有14个miRNA表达异常。其中，miR-1246是监管最为严格的。细胞分析表明miR-1246显著提高了A549肺癌细胞的迁移和侵袭能力。同时，免疫荧光分析显示miR-1246促进A549细胞EMT过程，同时E-cadherin表达降低，同时增加vimentin和转化生长因子β（TGF-β）的表达。此外，一个在线工具预测miR-1246可能与GSK-3β的3′未翻译区域结合，miR-1246检测的过度表达和敲除证实了这一点。

结论
综合研究表明miR-1246通过靶向GSK-3β/β-连环素调节Wnt/β-连环素途径，部分地促进肿瘤转移。MiR-1246在肺癌的诊断和治疗中可能起着重要作用。

Purpose
MicroRNAs (miRNAs) are a group of small non-coding RNAs involved in different cancers, including lung cancer. Here, we aim to investigate the expression profiles of circulating miRNAs and their roles contributed to the progress of lung cancer.


Materials and Methods
The levels of circulating miRNA in lung cancer patients were investigated by miRNAs assay. Then we predicted the target genes of aberrantly expressing miRNAs by searching genetic databases. Based on the A549 cells transfected with miR-1246 mimics or miR-1246 inhibitor,we further measured the roles of miR-1246 involving in the epithelial-mesenchymal transition (EMT), migration and invasion capacities of lung cancer cells in vitro. Finally, we detected the effects of miR-1246 on glycogen synthase kinase-3β (GSK-3β)/β-catenin pathway by immunofluorescence and Western blot, respectively.


Results
We identified that 14 miRNAs were aberrantly expressed in the serum of lung cancer patients. Among them, miR-1246 was the most up-regulated. The cell assays indicated that miR-1246 significantly increased the migration and invasion capabilities of A549 lung cancer cells. Meanwhile, immunofluorescence analysis revealed that miR-1246 promoted EMT process of A549 cells accompanying with decreasing E-cadherin expression, while increasing vimentin and transforming growth factor β (TGF-β) expression. Furthermore, an online tool predicated that miR-1246 might bind to 3′-untranslated region of GSK-3β, which was confirmed by overexpression and knockdown of miR-1246 assays.


Conclusion
Taken together, the study illustrates that miR-1246 regulates Wnt/β-catenin pathway through targeting GSK-3β/β-catenin, which partly contributing to tumor metastasis. MiR-1246 may play an essential role in the diagnosis and therapeutic of lung cancer.


https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6790833/