背景:microRNAs(miRNAs)参与肺腺癌(LUAD)的进展,然而,miR-6742-5p在LUAD中的功能仍然未知,从而进行了这项研究。
方法:从基因表达综合(GEO)数据库获得来自LUAD和正常对照的mRNA和miRNA表达数据,TargetScan和mirDIP用于预测miR-6742-5p和FGF8之间的关系。Q-PCR,westernblot,双荧光素酶,进行伤口愈合和transwell测定以测试miR-6742-5p在LUAD中的功能。
结果:生物信息学分析和双荧光素酶鉴定FGF8是miR-6742-5p的靶基因,在人类组织和细胞系的LUAD中下降,miR-6742-5POE抑制裸鼠LUAD的进展。MiR-6742-5pOE和KD抑制或增加通过伤口愈合和transwell测定H522和PC-9细胞测试的LUAD转移能力,这些关于miR-6742-5pOE的效应被FGF8逆转;miR-6742-5pOE,KD抑制并增加了FGF8的表达,作为其下游p-ERK1/2,MMP-2/-9,这些结果被ERK1/2抑制剂Ro67-7476纠正;miR-6742-5pKD增加了迁移和侵袭的细胞,并被MMPs抑制剂S3304抑制。这些结果鉴定了miR-6742-5p与FGF8-ERK1/2信号通路在LUAD进展中的负相关。
结论:我们得出结论,miR-6742-5p可能通过靶向FGF8/ERK1/2/MMPs信号通路成为LUAD进展的调节因子,这为LUAD提供了新的治疗靶点。
microRNAs (miRNAs) are involved in the progression of Lung adenocarcinoma (LUAD), however, the functions of miR-6742-5p in LUAD remains unknown, thereby this study was carried on.
The mRNA and miRNA expression data from the LUAD and normal control were obtained from Gene Expression Omnibus (GEO) database, TargetScan and mirDIP were applied to predict the relationship between miR-6742-5p and FGF8.Q-PCR, western blot, dual-luciferase, wound Healing and transwell assays were performed to test the functions of miR-6742-5p in LUAD.
Bioinformatics analysis and dual-luciferase identified FGF8 is the target-gene of miR-6742-5p, which is declined in LUAD of human tissues and cell lines, and miR-6742-5P OE suppressed the progression of LUAD in nude mice. MiR-6742-5p OE and KD suppressed or increased the abilities of LUAD\' metastasis tested by wound healing and transwell assays H522 and PC-9 cells, these effects about miR-6742-5p OE were reversed by FGF8; miR-6742-5p OE, KD inhibited and increased the expression of FGF8 as its downstream p-ERK1/2, MMP-2/-9, these results were corrected by ERK1/2 inhibitor: Ro 67-7476; the miR-6742-5p KD increased the migrated and invaded cells and suppressed by MMPs inhibitor: S3304. These results identified the negative correlation of miR-6742-5p with FGF8-ERK1/2 signal pathway in LUAD progression.
We conclude that miR-6742-5p might be a regulator of LUAD progression by targeting FGF8/ERK1/2/MMPs signaling pathway, which provides a novel therapeutic target for LUAD.