Serum MiR-4687-3p Has Potential for Diagnosis and Carcinogenesis in Non-small Cell Lung Cancer

Man Liu; Qiufang Si; Songyun Ouyang; Zhigang Zhou; Meng Wang; Chunling Zhao; Ting Yang; Yulin Wang; Xue Zhang; Wenbo Xie; Liping Dai; Jitian Li

doi:10.3389/fgene.2020.597508

Serum MiR-4687-3p Has Potential for Diagnosis and Carcinogenesis in Non-small Cell Lung Cancer

Front Genet. 2020 Nov 23:11:597508. doi: 10.3389/fgene.2020.597508. eCollection 2020.

Authors

Man Liu^{1

2}, Qiufang Si^{3

2}, Songyun Ouyang⁴, Zhigang Zhou⁵, Meng Wang⁵, Chunling Zhao⁴, Ting Yang^{3

2}, Yulin Wang^{1

2}, Xue Zhang^{1

2}, Wenbo Xie⁶, Liping Dai^{1

2}, Jitian Li⁷

Affiliations

¹ Henan Institute of Medical and Pharmaceutical Sciences, Academy of Medical Sciences, Zhengzhou University, Zhengzhou, China.
² Henan Key Laboratory of Tumor Epidemiology & State Key Laboratory of Esophageal Cancer Prevention, Zhengzhou University, Zhengzhou, China.
³ BGI College, Zhengzhou University, Zhengzhou, China.
⁴ Department of Respiratory and Sleep Medicine, The First Affiliated Hospital in Zhengzhou University, Zhengzhou, China.
⁵ Department of Radiology, The First Affiliated Hospital in Zhengzhou University, Zhengzhou, China.
⁶ Department of Computer Science, College of Engineering, University of Texas at El Paso, El Paso, TX, United States.
⁷ Laboratory of Molecular Biology, Henan Luoyang Orthopedic Hospital (Henan Provincial Orthopedic Hospital), Zhengzhou, China.

Abstract

The lack of a useful biomarker partly contributes to the increased mortality of non-small cell lung cancer (NSCLC). MiRNAs have become increasingly appreciated in diagnosis of NSCLC. In the present study, we used microarray to screen 2,549 miRNAs in serum samples from the training cohort (NSCLC, n = 10; the healthy, n = 10) to discover differentially expressed miRNAs (DEMs). Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) assay was applied to validate the expression level of selected overexpressed DEMs of NSCLC in a validation cohort (NSCLC, n = 30; the healthy, n = 30). Area under the receiver operating characteristic curve (AUC) was performed to evaluate diagnostic capability of the DEMs. The expression of the miRNAs in tissues was analyzed based on the TCGA database. Subsequently, the target genes of the miR-4687-3p were predicted by TargetScan. Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were tested by R software (ClusterProfiler package). NSCLC cells were transfected with inhibitor or mimic to down-regulate or up-regulate the miR-4687-3p level. The function of miR-4687-3p on proliferation, invasion, and migration of lung cancer cells were investigated through CCK-8 and Transwell assays, respectively. In the results, we identified serum miR-4687-3p that provided a high diagnostic accuracy of NSCLC (AUC = 0.679, 95%CI: 0.543-0.815) in the validation cohort. According to the TCGA database, we found that the miR-4687-3p level was significantly higher in NSCLC tissues than in normal lung tissues (p < 0.05). GO and KEGG pathway enrichment analysis showed that postsynaptic specialization and TGF-β signaling pathway were significantly enriched. Down-regulation of miR-4687-3p could suppress the proliferation, invasion, and migration of the NSCLC cells, compared with inhibitor negative control (NC). Meanwhile, overexpression of miR-4687-3p could promote the proliferation, invasion, and migration of the NSCLC cells compared with mimic NC. As a conclusion, our study first discovered that serum miR-4687-3p might have clinical potential as a non-invasive diagnostic biomarker for NSCLC and play an important role in the development of NSCLC.

Keywords: NSCLC; bioinformatics; biomarker; miR-4687-3p; microarray.