Hgf 1

All human oral cancer cell lines (Ca9-22, CAL 27, HSC-3, OC-2, and SCC-9) and a normal oral cell line (HGF-1) were used from Health Science Research Resources Bank (HSRRB) (Osaka, Japan) and American Type Culture Collection (ATCC; Manassas, VA, USA) except for OECM1 [57 (link)], a generous gift from Dr. Wan-Chi Tsai (Kaohsiung Medical University, Taiwan). Cells were cultured in 5% CO₂ at 37 °C with humidity and maintained by regular formula (Gibco, Grand Island, NY, USA) with 10% fetal bovine serum as previously described [49 (link)].
Manoalide (CAYMAN CHEMICAL, Ann Arbor, MI, USA) was dissolved in dimethyl sulfoxide (DMSO) for treatment. A ROS scavenger N-acetylcysteine (NAC) [58 (link)] (Sigma-Aldrich; St. Louis, MO, USA) was dissolved in double distilled water. The mitochondrial superoxide inhibitor MitoTEMPO [59 (link)] (Cayman Chemical, Ann Arbor, MI, USA), panapoptosis inhibitor Z-VAD-FMK [60 (link)], Cas 8 inhibitor Z-IETD-FMK, and Cas 9 inhibitor Z-LEHD-FMK (Selleckchem.com; Houston, TX, USA) was dissolved in DMSO. All experiments contain the same concentration of DMSO.

The human gingival squamous cell carcinoma (GSCC) cell line (Ca9-22) and gingival normal cell line (HGF-1) were obtained from Health Science Research Resources Bank (HSRRB, Osaka, Japan). The human GSCC OECM-1 cell line was purchased from EMD Millipore Corporation (Temecula, CA, USA). The normal human keratinocytes (HaCaT) cell line was provided by Prof. N.L. Wu (MacKay Memorial Hospital, Taipei, Taiwan). Cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum (FBS), penicillin (100 units/mL), and streptomycin (100 µg/mL) under humidified air containing 5% CO₂ at 37 °C. All cell culture reagents were purchased from Gibco-BRL life technologies (Grand Island, NY, USA).