Ox ldl

LncRNA CASC2 was amplified and then cloned into pcDNA3.1 vector (Invitrogen, US) to construct pcDNA-CASC2 overexpression plasmids. CASC2 siRNA (Si-CASC2), siRNA negative control (si-NC), PAPD5 siRNA (si-PAPD5) and PAPD5 si-NC, miR-532-3p mimic and miRNA mimic negative control (miR-NC) were purchased from GenePharma (Shanghai, China). The transfection was carried out by lipofectamine 2000 (Invitrogen, US). For observation of the effect of oxLDL (Biosynthesis, China) on the expression of CASC2 and miR-532-3p, VSMCs were cultured with 50 μg/ml ox-LDL for two d.

A human vascular smooth muscle cell (HA-VSMC) line was obtained from American Type Culture Collection (ATCC, Manassas, VA, USA), with 1% penicillin/streptomycin (Beyotime Biotechnology Company, Shanghai, China), cultured as previously described [19 (link)]. ox-LDL (Biosynthesis Biotechnology Company, Beijing, China) was used for AS model construction in vitro. To be specific, the cells were cultured in the medium with the presence of diverse dosages of ox-LDL (0 μg/mL, 25 μg/mL, 50 μg/mL, and 75 μg/mL) for 24 h and grown in the medium containing ox-LDL at a final concentration of 50 μg/mL for 24 h [11 (link)]. Short hairpin RNA (shRNA) targeting TUG1 (sh-TUG1), shRNA targeting ROR2 (sh-ROR2), TUG1 overexpression plasmid (TUG1), miR-141-3p inhibitor (anti-miR-141-3p), miR-141-3p mimic (miR-141-3p), and controls (sh-NC, pcDNA, anti-miR-NC, and miR-NC) were obtained from GenePharma (Shanghai, China). A Lipofectamine 3000 (Invitrogen, Carlsbad, CA, USA) kit was used for transfection according to the manufacturer's instructions. The sequences were shown as follows: sh-TUG1, sequence, 5′-GACTACCTTCCCTGTGCTATT-3′; sh-ROR2, sequence, 5′-GCCCGATTCCAACTCTGAAAG-3′.