miR-218-5p靶向EGLN3抑制肺腺癌增殖、迁移和侵袭

miR-218-5p inhibits proliferation, migration and invasion by lung adenocarcinoma cells by targeting EGLN3

目的 微小RNA(microRNAs, miRNA)参与了多种癌症的发生发展,有研究证明微小RNA-218-5p(microRNA-218-5p, miR-218-5p)在癌症的发展中发挥着重要作用。然而,miR-218-5p影响肺腺癌(lung adenocarcinoma, LUAD)发展的具体机制尚未明确。方法 通过癌症基因组图谱(the cancer genome atlas, TCGA)数据分析肺腺癌组织中miR-218-5p的表达情况,实时荧光定量PCR(quantitative real time polymerase chain reaction, qRT-PCR)检测miR-218-5p和EGLN3在人肺正常细胞系和人肺腺癌细胞系中的表达。蛋白质印迹法(Western blot)检测miR-218-5p对AKT/mTOR信号通路关键蛋白表达的影响以及细胞中EGLN3的蛋白表达。使用生物信息学方法预测并通过荧光素酶报告基因检测证实miR-218-5p与EGLN3的靶向关系。细胞活力和细胞毒性检测(cell counting kit-8, CCK-8)、细胞克隆形成、细胞划痕、Transwell实验检测miR-218-5p与EGLN3对肺腺癌细胞功能的影响。结果 miR-218-5p在肺腺癌中低表达,过表达miR-218-5p可抑制肺腺癌细胞的增殖、迁移与侵袭,并抑制AKT/mTOR信号通路的激活。miR-218-5p的下游靶基因EGLN3在肺腺癌中高表达。过表达EGLN3减弱了miR-218-5p过表达对肺腺癌细胞增殖、迁移和侵袭的抑制作用。结论 miR-218-5p靶向下调EGLN3抑制肺腺癌细胞的增殖、迁移与侵袭,这些结果为肺腺癌的新治疗方法和检测手段的开发提供了新的参考依据。

Objective MicroRNAs are involved in the occurrence and development of many cancers, and microRNA-218-5p(miR-218-5p) has been proven to play an important role in cancer development. However, the specific mechanism of miR-218-5p’s effect on the development of lung adenocarcinoma(LUAD) has not been clarified. Methods The expression of miR-218-5p in LUAD tissues was analyzed in TCGA data, and the mRNA expression of miR-218-5p and EGLN3 in human lung normal and human LUAD cell lines was detected by qRT-PCR. Western blot was used to detect the protein expression levels of key AKT/mTOR signaling pathway proteins and EGLN3 in tumor cells. The microRNA-target relationship between miR-218-5p and EGLN3 was predicted by bioinformatics method and confirmed by luciferase reporter gene detection. The roles of miR-218-5p and EGLN3 in LUAD were measured by CCK-8, colony formation, scratch healing, and Transwell assays. Results miR-218-5p was underexpressed in LUAD. Overexpression of miR-218-5p inhibited the proliferation, migration and invasion of LUAD cells and the activation of the AKT/mTOR signaling pathway. EGLN3 was the downstream target gene of miR-218-5p, which was highly expressed in LUAD. Overexpression of EGLN3 weakened the inhibitory effect of miR-218-5p on proliferation, migration and invasion by LUAD cells. Conclusions miR-218-5p inhibited proliferation, migration, and invasion in LUAD cells by negatively affecting EGLN3 expression.