Integration of bioinformatics analysis and experimental validation identifies plasma exosomal miR-103b/877-5p/29c-5p as diagnostic biomarkers for early lung adenocarcinoma.

Abstract

The aim of this study was to identify miRNAs in plasma exosomes as noninvasive biomarkers for the early diagnosis of lung adenocarcinoma (LUAD). First, exosomal miRNA profiling of three patients with early LUAD and three patients with benign lung disease were screened by next-generation sequencing (NGS) method. Sequencing results showed that 154 exosomal miRNAs were differentially expressed in the plasma of LUAD patients, among which 68 miRNAs were up-regulated and 86 miRNAs were down-regulated. GSE137140 is a GEO database containing serum miRNAs sequencing data from 1566 lung cancer patients and 1774 non-cancer patients controls. When comparing the sequencing data, it was found that most miRNAs (37/68) up-regulated in our LUAD group were also significantly up-regulated in GSE137140, suggesting that circulating miRNAs in lung cancer patients may be enriched in plasma exosomes. In GSE137140, the AUC of the combination of hsa-miR-103b, hsa-miR-29c-5p and hsa-miR-877-5p was 0.873, showing great potential as new tumor markers. To our knowledge, these three exosomal miRNAs have not been reported in lung cancer research. Furthermore, bioinformatics tools were used to analyze the target genes of three candidate miRNAs, which were indeed closely related to the occurrence and development of lung cancer. Bioinformatics algorithms deduced a highly conserved sequence in the 3'-UTR of SFRP4, FOXM1 and TMEM98 that could be bound with miR-103b/877-5p/29c-5p. A luciferase assay indicated that miR-103b/877-5p/29c-5p directly targeted the 3'-UTR of SFRP4, FOXM1 and TMEM98, respectively. Finally, three candidate miRNAs were validated by qRT-PCR in 17 early LUAD samples and 17 control plasma samples. Integration of bioinformatics analysis and experimental validation identifies, this study provides novel insights into miRNA-related networks in LUAD. Hsa-miR-103b, hsa-miR-29c-5p, and hsa-miR-877-5p may be used as diagnostic biomarkers for early LUAD.