AUTHOR=Wang Jin , Yu Xiao-fan , Ouyang Nan , Zhao Shiyu , Yao Haiping , Guan Xifei , Tong Jian , Chen Tao , Li Jian-xiang 

TITLE=MicroRNA and mRNA Interaction Network Regulates the Malignant Transformation of Human Bronchial Epithelial Cells Induced by Cigarette Smoke

JOURNAL=Frontiers in Oncology

VOLUME=Volume 9 - 2019

YEAR=2019

URL=https://www.frontiersin.org/journals/oncology/articles/10.3389/fonc.2019.01029

DOI=10.3389/fonc.2019.01029

ISSN=2234-943X

ABSTRACT=Objective: Aim of this study was to analyze the correlation and interaction of miRNAs and mRNAs regulatory network and their biological function during malignant transformation of BEAS-2B cells induced by cigarette smoke (CS).
Methods: Normal human bronchial epithelial cells (BEAS-2B) were continuously exposed to CS for 30 passages (S30) to establish an in vitro malignant transformation cell model.  The transformed cells were validated by scratch wound healing, transwell migration, colony formation and tumorigenicity assay. The miRNA and mRNA sequencing analysis was performed to identify differentially expressed miRNAs (DEMs) and differentially expressed genes (DEGs) between normal BEAS-2B and S30 cells. The miRNA-seq data of lung cancer with corresponding clinical data obtained from TCGA was used to further identify lung cancer-related DEMs and their correlations with smoking history. The target genes of these DEMs were predicted using the miRDB database, and their functions were analyzed using the online tool “Metascape”. 
Results: The migration ability, colony formation rate and tumorigenicity ability of S30 cells were enhanced. A total of 42 miRNAs and 755 mRNAs were dysregulated in S30 cells compared with normal BEAS-2B cells. The expression change of top five DEGs and DEMs were consistent with our sequencing results. Among these DEMs, eight miRNAs were found dysregulated in lung cancer tissues based on TCGA data. In these eight miRNAs, six of them including miR-96-5p, miR-93-5p, miR-106-5p, miR-190a-5p, miR-195-5p, and miR-1-3p, were found to be related to smoking history. Several DEGs, including THBS1, FN1, PIK3R1, CSF1, CORO2B, and PREX1, were involved in many biologic processes by enrichment analysis of miRNA and mRNA interaction. We identified the negatively regulated miRNA-mRNA pairs in the CS-induced lung cancer, which were implicated in several cancer-related (especially EMT-related) biological process and KEGG pathways in the malignant transformation progress of lung cells induced by CS.
Conclusion: Our result demonstrated the dysregulation of miRNA-mRNA profiles in cigarette smoke-induced malignant transformed cells, suggesting that several miRNAs may contribute to cigarette smoke-induced lung cancer. These genes may serve as biomarkers for predicting lung cancer pathogenesis and progression.  They can also be targets of novel anticancer drug development.