Atcc pcs 200 014

Human Tca8113, SCC-15 and CAL-27 TSCC cells, and normal gingival epithelial cells (ATCC^® PCS-200-014™) were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA). TSCC cells were maintained in Dulbecco's modified Eagle's medium supplemented with 10% heat-inactivated fetal bovine serum (FBS) (both from Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA), 100 U/ml penicillin and 100 mg/ml streptomycin. Normal gingival epithelial cells were cultured in minimum essential media (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS, 100 U/ml penicillin and 100 mg/ml streptomycin. Cells were maintained at 37°C in a humidified atmosphere containing 5% CO₂ and 95% air.

Overall, 4 OSCC cell lines (HSC2, HSC4, SAS and KON) were acquired from the Cell Bank of the Chinese Academy of Medical Sciences. The 4 OSCC cell lines were cultured in Dulbecco's modified Eagles medium F-12 HAM (Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) with 10% fetal bovine serum (FBS; Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA). Primary gingival keratinocytes (PGK; ATCC^® PCS-200-014™) were purchased from American Type Culture Collection (ATCC; Manassas, VA, USA) and cultured in Dermal Cell Basal Medium (ATCC^® PCS-200-030™) plus one Keratinocyte Growth Kit (ATCC^® PCS-200-040™).

Three human TSCC cell lines, SCC-9, CAL-27 and SCC-15, as well as normal gingival epithelial cells (ATCC^® PCS-200-014™) were purchased from the American Type Culture Collection (ATCC). Previous studies (26 (link),27 (link)) have used the normal gingival epithelial cells as a control for TSCC cell lines. Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin solution (all Invitrogen; Thermo Fisher Scientific, Inc.) was utilized for cell culture. All cells were maintained in a humidified incubator at 5% CO₂ and 37°C.

Normal gingival epithelial cells (ATCC^® PCS-200-014™) were acquired from the American Type Culture Collection (ATCC; Manassas, VA, USA) and cultured in minimum essential medium (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) containing 10% fetal bovine serum (FBS; Gibco; Thermo Fisher Scientific). Dulbecco’s modified Eagle’s medium (DMEM; Gibco; Thermo Fisher Scientific) with 10% FBS was employed in the culture of the TSCC cell line CAL-27 (ATCC). The other two TSCC cell lines SCC-9 and SCC-15 were maintained in a 1:1 mixture of DMEM and Ham’s F12 medium (Gibco; Thermo Fisher Scientific) containing 10% FBS and 400 ng/mL hydrocortisone. All cells were incubated at 37°C in a humidified incubator supplied with 5% CO₂.
Small interfering RNAs (siRNAs) for NOP14-AS1 (si-NOP14-AS1) and negative control (NC) siRNA (si-NC) were designed and offered by Shanghai GenePharma Co., Ltd. (Shanghai, China). The overexpressed high mobility group box 3 (HMGB3) plasmid pcDNA3.1-HMGB3 was constructed by RiboBio (Guangzhou, China). miRNA-665 (miR-665) mimic, NC mimic, miR-665 inhibitor, and NC inhibitor were also obtained from Shanghai GenePharma. TSCC cells were seeded into six-well plates, and transfection was performed by employing Lipofectamine 2000 (Invitrogen; Thermo Fisher Scientific).