This study tended to clarify the role of miR-126 in non-small cell lung cancer (NSCLC) cell biological behaviors in vitro, containing cell proliferation, migration, invasion, and apoptosis. miRNA expression microarray related to NSCLC was accessed from gene expression omnibus (GEO) database and subjected to differential analysis using the “limma” package. Real-time quantitative PCR was conducted to assess the expression of miR-126 in NSCLC cell lines. wIn vitro experiments including 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT), wound healing assay, Transwell, and flow cytometry assay were used for evaluating the effect of miR-126 on cell proliferation, migration, invasion, and apoptosis. Additionally, target mRNA for miR-126 was predicted and further validated by bioinformatics analysis and dual-luciferase reporter assay, respectively. It suggested that miR-126 was significantly down-regulated in NSCLS based on the expression microarray, and similar expression trend was exhibited in cancer cell lines. In the meantime, overexpression of miR-126 was found to result in inhibition of cell proliferation, migration, and invasion while promotion of cell apoptosis, with reductions in protein expression of AKT2 and phosphorylated HK2 (p-HK2) as well. AKT2, identified to be a direct target of miR-126 in NSCLC as judged by dual-luciferase reporter assay. Additionally, overexpression of AKT2 was observed to have the ability of elevating p-HK2 protein expression and reversing the effect of miR-126 on NSCLC cell proliferation, migration, and invasion. Given the above findings, we can see that miR-126 exerts its role in NSCLC cell proliferation, migration, invasion, and apoptosis with the aid of AKT2/HK2 axis.

中文翻译：

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miR-126通过AKT2/HK2轴调控非小细胞肺癌细胞的增殖、迁移、侵袭和凋亡

本研究旨在阐明miR-126在体外非小细胞肺癌（NSCLC）细胞生物学行为中的作用，包括细胞增殖、迁移、侵袭和凋亡。从基因表达综合 (GEO) 数据库访问与 NSCLC 相关的 miRNA 表达微阵列，并使用“limma”包进行差异分析。进行实时定量 PCR 以评估 miR-126 在 NSCLC 细胞系中的表达。w体外实验包括3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-溴化四唑(MTT)、伤口愈合试验、Transwell和流式细胞术试验用于评估效果miR-126 对细胞增殖、迁移、侵袭和凋亡的影响。此外，通过生物信息学分析和双荧光素酶报告分析预测并进一步验证了 miR-126 的靶 mRNA，分别。基于表达微阵列表明miR-126在NSCLS中显着下调，并且在癌细胞系中表现出相似的表达趋势。同时，发现 miR-126 的过表达会抑制细胞增殖、迁移和侵袭，同时促进细胞凋亡，同时减少 AKT2 和磷酸化 HK2 (p-HK2) 的蛋白表达。AKT2，经双荧光素酶报告基因测定确定为 NSCLC 中 miR-126 的直接靶标。此外，观察到 AKT2 的过表达具有提高 p-HK2 蛋白表达和逆转 miR-126 对 NSCLC 细胞增殖、迁移和侵袭的作用的能力。鉴于上述发现，我们可以看出miR-126在NSCLC细胞增殖、迁移、侵袭、