MicroRNA-144-3p Promotes Erastin-Induced Ferroptosis by Targeting SLC7A11 in NSCLC Cells

Abstract

Background: Non-small cell lung cancer (NSCLC) is the most frequent cause of malignancy-associated mortality worldwide. Ferroptosis is involved in tumorigenesis and metastasis of various cancers. Research showed that microRNA-144-3p (miR-144-3p) has regulatory impacts in NSCLC. However, the mechanism of this regulation is still unknown. Methods: Realtime quantitative polymerase chain reaction (RT-PCR) was applied to examine the level of miR-144-3p in NSCLC cells. MiR-144-3p overexpression plasmid was designed and constructed. Cell Counting Kit-8 (CCK-8) was conducted to assess the cell cytotoxicity. The malondialdehyde (MDA), reactive oxygen species (ROS), extracellular iron contents and glutathione (GSH) levels were estimated through enzyme-linked immunosorbent assay (ELISA) and flow cytometry. RNA (ribonucleic acid) pulldown and luciferase reporter assays were applied to assess the interaction between miR-144-3p and solute carrier family 7 member 11 (SLC7A11). Results: The study showed down-regulation of miR-144-3p in NSCLC clinical tissues and cells. MiR-144-3p overexpression significantly enhanced cytotoxicity induced by Erastin in NCI-H23 and NCI-H1650 cells. Further studies demonstrated that miR-144-3p overexpression accelerated the MDA, ROS and extracellular iron contents while inhibited GSH level in NCI-H23 cells treated with Erastin. Moreover, the luciferase reporter and RNA pulldown assay unveiled that SLC7A11 was a downstream target of miR-144-3p. Overexpression of SLC7A11 effectively reversed lipid peroxidation and cell cytotoxicity induced by miR-144-3p overexpression. Conclusions: To summarize, this study provided evidences on the functions and mechanism of ferroptosis activated by miR-144-3p and revealed that miR-144-3p might be a novel potential application for NSCLC patients.