miR-30d-5p represses the proliferation, migration, and invasion of lung squamous cell carcinoma via targeting DBF4

J Environ Sci Health C Toxicol Carcinog. 2021;39(3):251-268. doi: 10.1080/26896583.2021.1926855. Epub 2021 Jun 24.

Abstract

Objective: This study aims to explore the mechanism of miR-30d-5p in regulating the development of lung squamous cell carcinoma (LUSC) via targeting DBF4.

Methods: Bioinformatics methods were employed to analyze the differentially expressed genes in LUSC tissue microarray. qRT-PCR was employed to detect the expression of miR-30d-5p and DBF4 mRNA in normal human bronchial epithelial cells and LUSC cells. CCK-8 was used to detect LUSC cell activity. Wound healing assay was employed to detect the migratory ability of LUSC cells. Transwell was employed to detect invasive ability. Dual-luciferase reporter assay was used to detect the targeting relationship between miR-30d-5p and DBF4. Western blot was used to detect the protein expression of marker molecules associated with epithelial-mesenchymal transition (EMT).

Results: In this study, the expression of miR-30d-5p in LUSC cell lines was found to be obviously low compared with that in normal human bronchial epithelial cell line, which was opposite to the expression of DBF4. Dual-luciferase reporter assay verified that miR-30d-5p could target DBF4 and the overexpression of miR-30d-5p downregulated the expression of DBF4. Overexpression of DBF4 promoted the proliferation, migration, invasion, and EMT of LUSC, whereas over-expression of miR-30d-5p could weaken the promotion of DBF4 on cancer cells.

Conclusion: miR-30d-5p downregulates the expression of DBF4 to regulate the development of LUSC.

Keywords: DBF4; EMT; lung squamous cell carcinoma; miR-30d-5p; migration and invasion; proliferation.