Sw480 cell line

The hepatic cancer Huh-7 cell line, the pancreatic cancer Panc-1 cell line and the colorectal cancer SW480 cell line were purchased from the American Type Culture Collection. Huh-7, Panc-1 and SW480 cells were cultured in DMEM, RPMI-1640 and Leibovitz L-15 medium with 10% FCS, respectively. The construction, purification and infection of replication-deficient recombinant adenoviruses containing p53 (Ad-p53) or the bacterial lacZ gene (Ad-LacZ) have been described previously [11 (link)].

Luria broth was purchased from Amresco. ¹⁵NH₄Cl and D₂O were purchased from Cambridge Isotope Laboratories. Tranilast was purchased from Sigma. The SW-480 cell line was obtained from the American Type Culture Collection (CCL-288). The cDNA of S100A12 and the RAGE V domain were purchased from Mission Biotech Company using vectors pET21b for S100A12 and pET-32b (+) for the RAGE V domain. The genes were subcloned into Escherichia coli BL21 (DE3) (Novagen). The details of the purification process for obtaining the pure S100A12 and the RAGE V domain proteins are given in the Supporting Information. The cell proliferation reagent WST-1 (4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulfonate) was purchased from Roche. FPS-ZM1, an inhibitor of the RAGE V domain, was purchased from Calbiochem [46 (link)].

Human colon adenocarcinoma derived Caco2 cell line (well-differentiated) (G1–2) (from adenocarcinoma) and SW480 cell line (poorly-differentiated) (G3–4) (from adenocarcinoma grades III–IV) were purchased from the American Type Culture Collection (ATCC) Cell Bank (Manassas, Virginia). Cells were grown in Roswell Park Memorial Institute (RPMI) 1640 medium for Caco2 cells and Dulbecco’s Modified Eagle Medium (DMEM) for SW480 cells, supplemented with 10% fetal bovine serum (FBS), 2 mM glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin, in monolayer cultures, and incubated at 37 °C in a humidified atmosphere containing 5% CO₂ in air. The cells were allowed to grow for the established time and then harvested. For the experimental group, Autumn Royal and Egnatia GSEs were added to the medium at increasing concentrations, and the control group received no treatment; the cells were then incubated at 37 °C in a humidified 5% CO₂ incubator for the times required by the experiments indicated below.

R8-RGD peptides with a terminal cysteine modification [Cys-RRRRRRRR-c(RGDfK)] were purchased from Chinapeptides Co. Ltd. (Shanghai, China). PEI 25K was purchased from Sigma-aldrich and PEI 1.8K was purchased from Alfa Aesar. YOYO-1, Lyso-Tracker red, Hoechst 33342, Lipofectamine 2000 and Lipofectamine 3000 were purchased from Invitrogen (USA). pUNO1-hTRAILa (hTRAIL) and pUNO1-MCS (MCS) plasmid were obtained from In vivo Gen (San Diego, CA, USA) and purified with QIAGEN Plasmid Mega Kit (Qiagen GmbH, Hilden, Germany). Annexin V-FITC /PI apoptosis detection kit was purchased from Nanjing KeyGen Biotech. Co., Ltd (Nanjing,China). Antibodies used for western blotting were purchased from Cell Signaling Technology (CST, Beverly, MA, USA).
SW 480 cell line was obtained from American Type Culture Collection (ATCC, USA) and cultured in DMEM with 10% fetal bovine serum (Gibco), 100 units/mL penicillin G sodium and 100 μg/mL streptomycin sulfate and maintained at 37 °C in a humidified and 5% CO2 incubator.

SW480 cell line was obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). SW620 and its metastatic derivative SW620-LiM2 were obtained from Dr. Roger Gomis at IRB Barcelona [12 (link)], both lines were authenticated and KRAS mutation confirmed. NCM460 cell line [15 (link)] was a kind gift from Dr. Mary Pat Moyer (INCELL, San Antonio, TX, USA). All cells were grown in DMEM with 12.5 mM glucose, 4 mM glutamine, 5% Fetal Bovine Serum (10270, Gibco, ThermoFisher Scientific, Waltham, MA, USA) and 1% Streptomycin/penicillin at 37 °C in a 5% CO₂ atmosphere. Cell volume was determined using a Scepter^TM Handheld Automated Cell Counter (Merk Millipore, Burlington, MA, USA).