Si linc00461

Human normal breast epithelial cell line MCF 10A (CBP60419) and breast cancer cell lines AU565 (CBP60353), MCF-7 (CBP60380), MDA-MB-157 (CBP60381) and MDA-MB-231 (CBP60382) were all purchased from Cobioer (Nanjing, China). Plasmids including si-NC, si-LINC00461, inhibitor NC, miR-144-3p inhibitor, mimic NC, miR-144-3p mimic, oe-NC, oe-KPNA2 were ordered from GenePharma (Shanghai, China), and the siRNAs were sequenced as below: si-LINC00461: 5′-CTGCAAAGAAGCATAAAATGA-3′; si-NC: 5′-TTCTCCGAACGTG TCACGT-3′.
Before transfection, cells were grown in 6-well plates (3 × 10⁵ cells/well) until the density reached 50%. 250 μL of serum-free Opti-MEM (51985042, Gibco, Gaitherburg, MD, USA) was used to dilute the target plasmids (4 μg) and Lipofectamin 2000 reagent (10 μL; 11668-019, Invitrogen, NY, California, USA), respectively. Thereafter, the dilutions were mixed, and then dripped into cells after 20 min. All cells were maintained in 5% CO₂ at 37 °C for 6 h, and cultured for additional 36–48 h with fresh mediums.

Human DLBCL cell lines, OCI-Ly7, GM12878, FARAGE, U2932, and TMD8, were maintained in the DMEM complete medium containing 10% fetal bovine serum (FBS), 100 mg of streptomycin/mL, and 100 units of penicillin/mL (Solarbio, Beijing, China). The cells were cultured in a 5% CO₂ incubator at 37°C and 95% humidity.
To analyze the influence of LINC00461, miR-411-5p, and BNIP3 on the biological behavior of DLBCL cells, the pcDNA3.1 carried human full-length LINC00461, siLINC00461, BNIP3, miR-411-5p mimic, and corresponding negative control (NC) plasmids were from GenePharma (Shanghai, China). After the cells in the 6-well plate reached 60%, Opti (100 μL, Invitrogen, Waltham, USA) and Lipofectamine^TM 2000 (5 μL, Invitrogen, Waltham, USA) were added and incubated for 5 min (A). Opti (100 μL), pcDNA3.1 DNA (20 ng/μL), mimic, or NC were added and incubated for 5 min (B). A and B were mixed and incubated for 20 min. After 16 h, the medium was changed, and the cells were harvested for subsequent experiments.