Investigating miR-152-3p Regulation of lncRNA H19 in Lung Cells

Lung epithelial cells (BEAS-2B; ATCC) were cultured at 37˚C in a 5% CO₂ incubator with a culture medium system containing Dulbecco's modified Eagle's Medium (DMEM) (Hyclone; Cytiva), 10% fetal bovine serum (FBS; Gibco; Thermo Fisher Sceintific, Inc.), and 1% penicillin/streptomycin solution (100X; Solarbio Life Science). starBase2.0(26 (link)) predicted the targeting binding loci between lncRNA H19 and miR-152-3p. GLO-H19-wild type (wt) and GLO-H19-mutant (mut) vectors were constructed and co-transfected with miR-152-3p mimics (5'-UCAGUGCAUGACAGAACUUGG-3') and NC-mimics (5'-ACUACUGAGUGACAGUAGA-3') (designed and synthesized by Sangon Biotech Co., Ltd.) respectively. The transfection kit used was Lipofectamine 2000. The transfected cells were cultured on 96-well plates for 48 h. Then, a Dual-luciferase reporter gene assay system (Promega Corporation) was employed to determine the luciferase activity. In this study, pGLO plasmid vectors (Bio-Rad Laboratories, Inc.) were used. The relative luciferase activity was measured as follows: Relative luciferase activity=luciferase activity of glowworm/internal reference luciferase activity of sea pansy.