Sulfasalazine (Azulfidine) is a synthetic salicylic acid derivative with affinity for elastin-containing connective tissues and formulated as a prodrug. Sulfasalazine induces iron death and inhibits NF-κB, TGF-β and COX-2.

METHODS: Rat glioma cells F98 and human glioma cells U251 were treated with Sulfasalazine (200-400 µM) for 96 h. Cell viability was measured by MTT assay.
RESULTS: Cell viability was significantly reduced in F98 at 200-400 µM Sulfasalazine concentration and U251 showed reduced cell viability at 400 µM Sulfasalazine. [1]
METHODS: Mouse melanoma cells B16F10 and mouse embryonic fibroblasts MEF were treated with Sulfasalazine (10-1000 µM) for 24 h. Cellular ROS levels were measured by DCFDA staining.
RESULTS: At lower Sulfasalazine concentrations (10-100 µM), no increase in intracellular ROS was observed. At higher concentrations of Sulfasalazine (800-1000 µM), there was an approximately 2.3-fold increase in intracellular ROS in B16F10 cells, while no increase in ROS was observed in MEF cells. [2]

METHODS: To detect antitumor activity in vivo, Sulfasalazine (250 mg/kg) was administered intraperitoneally to C57BL/6N mice bearing B16F10 xenografts once daily for three days. Twenty-four hours after the third Sulfasalazine dose, local X-ray irradiation was applied to anesthetized tumor-bearing C57BL/6N mice at a dose of 4 Gy.
RESULTS: Sulfasalazine alone did not significantly inhibit tumor growth; X-ray irradiation partially reduced tumor growth; the combination of Sulfasalazine and X-rays synergistically reduced tumor growth. [2]

Sulfasalazine is dissolved in culture medium. SW620 cells are grown in Dulbecco's modified Eagle medium, supplemented with 10% heat-inactivated FCS, 2 mmol/liter glutamine, and 1% (wt/vol) penicillin/streptomycin. SW620 cells are transfected with the 3xIgkBLuc reporter construct. After 18 h, cells are incubated with either medium alone or with sulfasalazine (0.1, 0.2, 0.5, 1, 2, 5 mM) before stimulation with TNFα, LPS, or PMA. Luciferase assay is performed[1].