目的：探讨miR-93/Eph受体A4（EphA4）分子轴通过细胞外调节蛋白激酶（extracellular regulated protein kinases，ERK）通路对非小细胞肺癌（non-small cell lung cancer，NSCLC）H460 和H1299 细胞增殖和迁移的影响。方法：用qPCR 检测H460 和H1299 细胞中miR-93 表达水平。分别在H460 细胞中转染miR-93 模拟物（mimics）和EphA4 过表达质粒、在H1299 细胞中转染miR-93 抑制剂（inhibitor）后，用MTT、Transwell 实验检测miR-93 对转染细胞增殖和迁移的影响。用双荧光素酶报告基因实验验证miR-93 与EphA4 之间的靶向调控关系。用Western blotting检测细胞中增殖细胞核抗原（proliferating cell nuclear antigen，PCNA）、EphA4、ERK和p-ERK 蛋白的表达水平，用MTT、Transwell 实验检测同时过表达miR-93 和EphA4 对H460 细胞增殖和迁移的影响。结果：miR-93 在H1299 细胞中表达水平高于H460 细胞（P<0.01）。过表达miR-93 促进H460 细胞增殖和迁移（均P<0.01），敲低miR-93 抑制H1299 细胞增殖和迁移（均P<0.01）。双荧光素酶报告基因实验证实miR-93靶向调控EphA4，过表达miR-93明显下调H460细胞中EphA4 mRNA和蛋白表达水平（均P<0.05），过表达miR-93 通过靶向EphA4 并激活ERK通路促进H460 细胞的增殖和迁移（均P<0.01）。结论：miR-93 促进NSCLC细胞增殖和迁移，其机制可能与靶向调控EphA4 并激活ERK通路有关。

Objective: To investigate the effect of miR-93/EphA4 (Eph receptor A4) axis on the proliferation and migration of nonsmall cell lung cancer (NSCLC) H460 and H1299 cells via regulating extracellular regulated protein kinases (ERK) pathway. Methods:The expression levels of miR-93 in H460 and H1299 cells was detected by qPCR. miR-93 mimics and EphA4 overexpression plasmids were transfected into H460 cells and miR-93 inhibitor was transfected into H1299 cells respectively, after which MTT assay and Transwell assay were used to detect the effects of miR-93 on proliferation and migration of transfected cells. The targeted regulatory relationship between miR-93 and EphA4 was verified by Dual-luciferase reporter gene assay. The expression levels of PCNA (proliferating cell nuclear antigen), EphA4, ERK and p-ERK were detected byWestern blotting. The effects of simultaneous overexpression of miR-93 and EphA4 on proliferation and migration of H460 cells were detected by MTT assay and Transwell assay. Results: The expression of miR-93 in H1299 cells was higher than that in H460 cells (P<0.01). Overexpression of miR-93 promoted proliferation and migration of H460 cells (all P<0.01), and knockdown of miR-93 inhibited proliferation and migration of H1299 cells (all P<0.01). The Dualluciferase reporter gene assay confirmed that miR-93 could target EphA4. Overexpression of miR-93 down-regulated the mRNA and protein expression levels of EphA4 (all P<0.05), and promoted proliferation and migration of H460 cells through targeted regulation of EphA4 and activation of ERK pathway (all P<0.01). Conclusion: miR-93 promotes the proliferation and migration of NSCLC cells,and its mechanism may be related to the targeted regulation of EphA4 and activation of the ERK pathway.

国家自然科学基金资助项目（No.81672833）