Department of Respiratory and Critical Medicine, Affiliated Taizhou Hospital of Zhejiang Province of Wenzhou Medical University, Taizhou, China.
Eur Rev Med Pharmacol Sci. 2020 Jan;24(2):721-727. doi: 10.26355/eurrev_202001_20051.
To explore the effects of micro ribonucleic acid (miR)-188 on proliferation and apoptosis of lung adenocarcinoma (LUAD) cells, and its potential mechanism.
The expression level of miR-188 in LUAD cell lines was detected via quantitative Real Time-Polymerase Chain Reaction (PCR). The effects of miR-188 overexpression on proliferation and apoptosis of A549 cells were detected using methyl thiazolyl tetrazolium (MTT) assay, colony formation assay, and flow cytometry. The potential targets for miR-188 were predicted using the TargetScan Human database, and the interaction between miR-188 and target gene was determined through Dual-Luciferase reporter assay. Moreover, the associations of miR-188 and sine oculis homeobox homolog 1 (SIX1) with the extracellular signal-regulated kinase (ERK) pathway were detected via Western blotting.
The expression of miR-188 significantly declined in LUAD cell lines (p<0.05). The overexpression of miR-188 significantly reduced the proliferation rate of A549 cells and increased the percentage of apoptotic A549 cells (p<0.05). Similarly, it was found in colony formation assay that the overexpression of miR-188 inhibited the colony formation ability of A549 cells most significantly (p<0.05). SIX1 was a direct target for miR-188, and its mRNA and protein expressions were downregulated by the overexpression of miR-188. The remarkable downregulation of phosphorylated ERK was observed in A549 cells overexpressing miR-188, while the decline in phosphorylated ERK was reversed in A549 cells overexpressing miR-188 and SIX1.
The expression of miR-188 is downregulated in LUAD cell lines. The overexpression of miR-188 inhibits proliferation and promotes apoptosis of LUAD cells, whose functional mechanism may be related to its regulation on the ERK signaling pathway by targeting SIX1.
探讨微小 RNA-188（miR-188）对肺腺癌（LUAD）细胞增殖和凋亡的影响及其潜在机制。
采用实时定量聚合酶链反应（qRT-PCR）检测 LUAD 细胞系中 miR-188 的表达水平。采用噻唑蓝（MTT）比色法、集落形成实验和流式细胞术检测 miR-188 过表达对 A549 细胞增殖和凋亡的影响。利用 TargetScan Human 数据库预测 miR-188 的潜在靶基因，并通过双荧光素酶报告基因实验验证 miR-188 与靶基因的相互作用。此外，通过 Western blot 检测 miR-188 与 sine oculis homeobox homolog 1（SIX1）与细胞外信号调节激酶（ERK）通路的相关性。
miR-188 在 LUAD 细胞系中的表达显著下调（p<0.05）。miR-188 过表达显著降低了 A549 细胞的增殖率，并增加了凋亡 A549 细胞的比例（p<0.05）。同样，在集落形成实验中，miR-188 的过表达最显著地抑制了 A549 细胞的集落形成能力（p<0.05）。SIX1 是 miR-188 的直接靶基因，miR-188 的过表达下调了其 mRNA 和蛋白表达。在过表达 miR-188 的 A549 细胞中观察到磷酸化 ERK 的显著下调，而在过表达 miR-188 和 SIX1 的 A549 细胞中，磷酸化 ERK 的下降得到逆转。
miR-188 在 LUAD 细胞系中表达下调。miR-188 的过表达抑制 LUAD 细胞的增殖并促进其凋亡，其功能机制可能与其通过靶向 SIX1 调节 ERK 信号通路有关。
