Mir 149 mimic

Human NSCLC cell lines (A549, H460 and H1299) and the normal human bronchial epithelial cell line 16HBE were obtained from the American Type Culture Collection and were cultured in DMEM (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) at 37°C with 5% CO₂.
miR-149 mimic, pre-negative control (NC), miR-149 inhibitor, NC, small interfering (si)RNA targeting BCYRN1 (si-BCYRN1) or targeting PKM2 (si-PKM2), and si-NC were synthesized by Guangzhou RiboBio Co., Ltd. The corresponding sequences were as follows: miR-149 mimic, 5′-UCUGGCUCCGUGUCUUCACUCCC-3′; pre-NC, 5′-UUCUCCGAACGUGUCACGU-3′; miR-149 inhibitor, 5′-GGGAGUGAAGACACGGAGCCAGA-3′; NC, 5′-CAGUACUUUUGUGUAGUACAA-3′; si-BCYRN1, 5′-CUCCAGAAAAAGGAAAAAAAAAA-3′; si-PKM2, 5′-GGAAAGAACAUCAAGAUUAUC-3′; si-NC, 5′-UUCUCCGAACGUGUCACGU-3′. Plasmid pcDNA-BCYRN1 or pcDNA-PKM2 were constructed by Shanghai GenePharma Co., Ltd. for the overexpression of BCYRN1 or PKM2 in cells. A459 or H1299 cells (2×10⁵) were transfected with 50 nM plasmid or 50 nM RNAs using Lipofectamine^® 3000 (Invitrogen; Thermo Fisher Scientific, Inc.), according to the manufacturer's protocol. After a 48-h incubation at 37°C, cells were harvested for subsequent experiments. An empty plasmid was used as the negative control.