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.

Comprehensive Analysis of Circular RNA Expression Profiles in Gefitinib-Resistant Lung Adenocarcinoma Patients

“…The RNA networks of circRNA-miRNA are presented in Figure 6 . In particular, we found that hsa_circ_0030591 (associated with the ABC transporters pathway) has 12 predictive target miRNAs related to lung cancer with one or more than one binding site, including hsa-let-7, 12 hsa-miR-103b, 13 hsa-miR-125a-5p, 14 hsa-miR-23, 15 and hsa-miR-449c-5p. 16 Similarly, hsa_circ_0040348 (associated with the PI3K-Akt signaling pathway) has 5 corresponding miRNAs with more than one binding site, and was predicted to bind to many lung cancer-related miRNAs with binding site, such as hsa-let-7, 12 hsa-miR-27a-3p, 17 hsa-miR-432-5p, 18 hsa-miR-513c-5p, 19 hsa-miR-622, 20 and hsa-miR-548.…”

Diagnostic value of exosomal noncoding RNA in lung cancer: a meta-analysis

“…Most of the articles provided and specificity information, which could be obtained directly, and the sensitivity and specificity of the other 9 articles were obtained by reading ROC curves with the digitizing software engage Digitizer ( 18 , 23 – 25 , 36 , 41 , 52 – 54 ). The results are shown in Table 1 and Supplementary Table 2 .…”

Elevated miR-29c-5p Expression in Nipple Aspirate Fluid Is Associated with Extremely High Mammographic Breast Density

“…Or, this miRNA could be selectively released into NAF leading to differing cellular and extracellular miRNA profiles. Interestingly, the miR-29 family has been reported to frequently reside in extracellular exosomes and upregulation of plasma exosomal miR-29c-5p has recently been suggested as a diagnostic biomarker for early lung carcinoma [ 41 ]. Furthermore, miR-29c-5p was upregulated in bladder serum cancer samples compared to normal samples and its dysregulation was correlated to advanced stage and poor outcome in bladder cancer patients [ 42 ].…”