The Second People's Hospital of China Three Gorges University, Yichang, China.
Hubei Key Laboratory of Tumor Microenvironment and Immunotherapy, Yichang, China.
Cancer Res Treat. 2019 Oct;51(4):1420-1429. doi: 10.4143/crt.2018.638. Epub 2019 Mar 4.
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.
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.
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.
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.
微小 RNA（miRNA）是一组参与包括肺癌在内的不同癌症的小非编码 RNA。在这里，我们旨在研究循环 miRNA 的表达谱及其对肺癌进展的作用。
通过 miRNA 检测研究肺癌患者循环 miRNA 的水平。然后，我们通过搜索遗传数据库预测异常表达 miRNA 的靶基因。基于 miR-1246 模拟物或 miR-1246 抑制剂转染的 A549 细胞，我们进一步测量了 miR-1246 涉及体外肺癌细胞上皮-间充质转化（EMT）、迁移和侵袭能力的作用。最后，我们通过免疫荧光和 Western blot 分别检测了 miR-1246 对糖原合酶激酶-3β（GSK-3β）/β-连环蛋白通路的影响。
我们发现 14 种 miRNA 在肺癌患者的血清中异常表达。其中，miR-1246 表达上调最明显。细胞实验表明，miR-1246 显著增加了 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，伴随 E-钙黏蛋白表达降低，而 vimentin 和转化生长因子β（TGF-β）表达增加。此外，在线工具预测 miR-1246 可能结合 GSK-3β 的 3'-非翻译区，这通过 miR-1246 的过表达和敲低实验得到证实。
综上所述，该研究表明 miR-1246 通过靶向 GSK-3β/β-catenin 调节 Wnt/β-catenin 通路，部分促进肿瘤转移。miR-1246 可能在肺癌的诊断和治疗中发挥重要作用。
