miR-21 Exerts Anti-proliferative and Pro-apoptotic Effects in LPS-induced WI-38 Cells via Directly Targeting TIMP3

• Published in: Cell biochemistry and biophysics 2021-12, Vol.79 (4), p.781-790
• Main Authors: Li, Jin-Xiu, Li, You, Xia, Tian, Rong, Feng-Yan
• Format: Article
• Language: English
• Subjects: Antagomirs - metabolism; Apoptosis; Apoptosis - drug effects; BAX protein; Bcl-2 protein; Binding sites; Biochemistry; Biological and Medical Physics; Biomedical and Life Sciences; Biophysics; Biotechnology; Cancer; Caspase-3; Caspase-9; Cell Biology; Cell Line; Cell Proliferation - drug effects; Diploids; Embryo fibroblasts; Enzyme-linked immunosorbent assay; Fibroblasts; Fibroblasts - cytology; Fibroblasts - drug effects; Fibroblasts - metabolism; Fibrosis; Gene expression; Hospitals; Humans; Idiopathic Pulmonary Fibrosis - chemically induced; Idiopathic Pulmonary Fibrosis - metabolism; Idiopathic Pulmonary Fibrosis - pathology; Inflammation; Interleukin 10; Interleukin 6; Interleukin-6 - genetics; Interleukin-6 - metabolism; Life Sciences; Lipopolysaccharides; Lipopolysaccharides - pharmacology; Lung diseases; MicroRNAs - genetics; MicroRNAs - metabolism; miRNA; Original Paper; Pharmacology/Toxicology; Pharmacy; Pneumonia; Pulmonary fibrosis; Reagents; Tissue inhibitor of metalloproteinase 3; Tissue Inhibitor of Metalloproteinase-3 - genetics; Tissue Inhibitor of Metalloproteinase-3 - metabolism; Western blotting
• ISSN: 1085-9195; 1559-0283; 1559-0283
• DOI: 10.1007/s12013-021-00987-w

Idiopathic pulmonary fibrosis (IPF) is a progressive lung disease, which was caused by a complex interplay of inflammatory responses and chronic damage. miR-21 is increased in patients with IPF, but its function in the embryonic lung-derived diploid fibroblasts cells subjected to LPS is elusive. miRNA expression profile was obtained from GEO database and target genes of miRNAs were forecasted by TargetScan. To mimic the LPS-induced injury, different concentrations of LPS were applied to treat WI-38 cells. Functional in vitro experiments were conducted to examine the role of miR-21 and TIMP3. Luciferase report assay was performed to verify the relationship between miR-21 and TIMP3. qRT-PCR, western blotting, and ELISA were conducted to detect the levels of the related miRNAs, proteins, and inflammatory factors. miR-21 presented higher levels in interstitial pneumonia patients and LPS-induced WI-38 cells. Overexpression of miR-21 was negatively correlated with the proliferative capability of LPS-treated WI-38 cells. miR-21 directly targets TIMP3. TIMP3 restored the suppressive impact of miR-21 mimic on the proliferation, while TIMP3 alleviated the promoting impact of miR-21 mimic on the apoptosis of WI-38 cells treated by LPS. miR-21 inhibited Bcl-2 but increased Bax, cleaved caspase-3, and cleaved caspase-9. Besides, miR-21 elevated the levels of IL-6 and IL-β but reduced the IL-10, which were weakened by TIMP3. Totally, miR-21 aggravated the LPS-induced lung injury and modulated inflammatory responses by targeting TIMP3.