Ac lehd cho

The human MPNST cell line FMS-1 [23] (link) and the human UPS cell line FPS-1 [22] (link) used in the present study, were established in our laboratory., Both FMS-1 and FPS-1 cells showed overexpression of COX-2, as demonstrated by immunohistochemistry, Western blotting, and reverse transcription-polymerase chain reaction analysis [51] (link), [52] . Cells were grown in RPMI-1640 medium (Sigma R8758, St. Louis, MO, USA) supplemented with 15% heat-inactivated fetal calf serum (FCS) (JRH Biosciences, Lenexa, KS, USA), 50 units/ml penicillin G, and 50 µg/ml streptomycin. The cells were inoculated into 25-cm² tissue culture flasks (Iwaki Glass, Tokyo, Japan) and incubated at 37°C in a humidified atmosphere with 5% CO₂.
Etodolac, a selective COX-2 inhibitor, was provided by Nippon Shinyaku Co. (Kyoto, Japan). A stock solution of etodolac was prepared in 99.5% ethanol and stored at −80°C. Caspase inhibitors (Z-VAD-FMK, broad caspase inhibitor; Ac-IETD-CHO, caspase-8 inhibitor; Ac-LEHD-CHO, caspase-9 inhibitor; Ac-DMQD-CHO, caspase-3 inhibitor) were obtained from Peptide Institute, Inc. (Osaka, Japan) and were dissolved in dimethyl sulfoxide (DMSO) as stock solutions.

Edible plant oils (canola oil, corn oil, extra virgin olive oil, grape seed oil, linseed oil, perilla oil, rice bran oil, safflower oil, sesame oil (non-roasted), sesame oil (roasted) and soybean oil) were obtained from a local supermarket (Aichi, Japan). N-acetyl-cysteine (NAC) was obtained from Sigma-Aldrich (St. Louis, MO, USA). Caspase-3 inhibitor (Ac-DNLD-CHO), caspase-9 inhibitor (Ac-LEHD-CHO), and pan-caspase inhibitor (Z-VAD-FMK) were purchased from Peptide Institute (Osaka, Japan). Vitamin E was obtained from Fujifilm Wako Pure Chemical Corporation (Osaka, Japan). Anti-AIF antibody was purchased from Proteintech (Rosemont, IL, USA). Anti-Bax, anti-Bcl-2, anti-caspase-3, anti-COX IV, anti-cytochrome c, and anti-poly (ADP-ribose) polymerase (PARP) antibodies, as well as a secondary horseradish peroxidase-conjugated anti-rabbit antibody, were obtained from Cell Signaling Technology (Danvers, MA, USA).

Phorbol 12-myristate 13-acetate (PMA), PI, caffeine, and DMSO were purchased from Sigma-Aldrich (St. Louis, MO, USA). Z-VAD-fmk, Ac-LEHD-cho, Ac-IETD-cho, and Ac-DNLD-cho were purchased from Peptide Institute, Inc. (Osaka, Japan). Cleaved caspase-3 antibody (#9661), caspase-8 antibody (#9746), caspase-9 antibody (#9502), PARP (46D11) rabbit monoclonal antibody (#9532), β-actin antibody (#4967), anti-rabbit IgG horseradish peroxidase (HRP)-linked antibody (#7074), anti-mouse IgG HRP-linked antibody (#7076), phosphor-ATM (Ser1981) (D25E5) antibody (#13050), and Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (#4412) were purchased from Cell Signaling Technology Japan, K.K. (Tokyo, Japan). The fluorescein isothiocyanate (FITC)-labeled monoclonal antibody anti-human cluster of differentiation 95 (CD95-FITC) was purchased from BioLegend (San Diego, CA, USA). FITC-conjugated mouse IgG₁ antibody was purchased from Beckman–Coulter (Fullerton, CA, USA).