Parthenolide

Pilocarpine and methoctramine were purchased from Sigma-Aldrich (St. Louis, MO, USA). Parthenolide, LY294002 and U0126 were purchase from Selleck (Shanghai, China). Antibodies used for Western blotting were purchased from Cell Signaling Technology (Danvers, MA, USA) for Erk, phospho-p42/44 Erk (Thr202/ Tyr204), Akt, phospho-Akt (Ser473), E-cadherin, vimentin, MMP9, Snail, Zeb1, phospho-NF-κB p65 (Ser536), NF-κB p65, IκBα, phosphor-IκBα (Ser32)and β-actin.

Female athymic BALB/c nude mice (6–8 weeks old) were purchased from Shanghai Experimental Animal Center, Chinese Academy of Science. The nude mice were subcutaneously injected with 5×10⁶ SGC7901 cells suspended in PBS (phosphate buffer saline) and grew until the tumors reached ~ 200 mm³. These mice were randomly divided into five groups and treated with LPS (lipopolysaccharide, 1 mg/kg/3d, enterocoelia), BA (Betulinic acid, 20 mg/kg/3d, intragastric), miRNA mimics (10 μg/week/mouse, tumor), gastrin (2 mg/kg, twice/day, subcutaneous), and PBS once daily (n = 6 for each group) for 14 days. These mice were also randomly divided into four groups and treated with ERK inhibitor PD98059 (10 mg/kg/day, once per 3 days, Selleck), P65 inhibitor PN (Parthenolide, 4 mg/kg/day, once per day, Selleck), miR23a/27a/24 inhibitors (10 μg/week/mouse, GenePharma), and the vehicle control for 14 days. All mice were sacrificed after anesthesia and tumor size and weight were measured. Tumor volume was calculated, V = length×width²/2 mm³. The experiments were approved by the animal research committee in Shanghai Jiao Tong University.