Tscca

The four TSCC cell lines TSCCA, TCA8113, CAL-27, and SCC-25 were obtained from Procell Life Science & Technology (Wuhan, China). TSCCA and TCA8113 cells were cultured in RMPI 1640 (Gibco, USA) plus 10% fetal bovine serum (FBS, Hyclone, USA). CAL-27 and SCC-25 were cultured in Dulbecco’s modified Eagle’s medium (DMEM, Gibco, USA) plus 10% FBS and DMEM/F12 (Gibco, USA) plus 10% FBS, respectively. All cell lines were incubated in a cell incubator at 37°C in 5% CO₂.
MiR-532-3p mimics/NC mimics and miR-532-3p inhibitor/NC inhibitor were transfected in CAL-27 and TCA8113 using Lipofectamine 2000 (Invitrogen, USA), respectively. MiR-532-3p mimics and CCR7/vector were co-transfected in CAL-27 using Lipofectamine 2000. The co-transfected cells were cultured in complete medium (DMEM+10% FBS+350 ng/ml CCL21). In addition, the CAL-27 cells were divided into two groups, Control and CCL21 (medium contains 350 ng/ml CCL21).

Normal Human Oral Keratinocyte (HOK) cell lines were obtained from TongPai (Shanghai) Biotechnology Co., Ltd (Shanghai, China). Human HNSCC cell lines Cal-27, FADU, and TSCCA were obtained from Procell Life Science & Technology Co., Ltd (WuHan, China). HOK cell lines were cultured in Minimum Essential Medium (MEM, HyClone, USA) with 10% fetal bovine serum (FBS) supplements (Gibco, New Zealand) and 1% penicillin plus streptomycin (BI, Israel) at 37°C in 5% CO₂. Cal-27, FADU, and TSCCA cell lines were seeded in Dulbecco’s modified Eagle’s medium (DMEM, HyClone, USA) with 10% FBS (Gibco, New Zealand) and 1% penicillin plus streptomycin (BI, Israel). The cells were incubated at 37°C in a 5% CO₂ atmosphere. Screening of cell lines with the highest lncRNA FOXD2-AS1 expression levels was carried out via RT-qPCR. The cells were seeded in 6-well plates, grown to 50–70% confluency, and infected with lentiviruses for lncRNA FOXD2-AS1 knockdown expression (sh-FOXD2-AS1), and knockdown expression negative control, (sh-NC) (GENECHEM, Shanghai, China). Transfection efficiency was analyzed after 72 hours.