Mucin type 2 mucin from porcine stomach

Bullfrog oil was provided by Asmarana Produtos Naturais (Natal, Brazil). Sodium hydroxide (NaOH), dimethyl sulfoxide (DMSO), potassium hydroxide, diethyl ether, hydrochloric acid (HCl), Tween^® 20, sodium thiosulfate, and sodium carbonate were from VETEC (Rio de Janeiro, Brazil). Miglyol^® 812 was a gift from Sasol (Witen, Germany). Butylhyldroxytoluene (BHT), buthylhydroxyanisole, potassium biphthalate, potassium dichromate, and Wijs solution were purchased from Labsynth (São Paulo, Brazil). Acetic acid, ethanol P.A, sodium bicarbonate, potassium iodide, starch, and chloroform were from Isofar (Rio de Janeiro, Brazil). Sucralose, Tutti-frutti flavoring and Acesulfame K were purchased from Valdequímica (São Paulo, Brazil). Xanthan gum, sodium benzoate and propylparaben were from ViaFarma (São Paulo, Brazil). Phenolphthalein was provided from Biotec Chemicals (Londrina, Brazil). Mucin Type II (Mucin from porcine stomach) and Span^® 80 were from Sigma-Aldrich (São Paulo, Brazil).

Briefly, 5-fluorouracil (Nanjing Wellchem Enterprise, Nanjing, China), chondroitin sulfate (Summit Nutritionals International™, Lebanon, NJ, USA), sodium deoxycholate (Sigma-Aldrich, São Paulo, Brazil), hydroxypropyl-methyl-cellulose (Methocel™ F4M—Sigma-Aldrich, São Paulo, Brazil), mucin Type II (Mucin from porcine stomach, Sigma-Aldrich, São Paulo, Brazil), sulfuric acid (Vetec Química Fina Ltda, Rio de Janeiro, Brazil), and methanol (Merck, Rio de Janeiro, Brazil, high-performance liquid chromatographic-grade) were used in these studies. MTT reagent (3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide) (Sigma Aldrich Inc., St. Louis, MO, USA). Epithelial human melanoma cell lines (A2058 (ATCC^® CRL-11147™) and A375 (ATCC^® CRL-1619™ All)), donated by prof. Dr. José Alexandre Barbuto—USP Institute of Biomedical Sciences, Tumor Immunology Laboratory (São Paulo, Brazil). All chemicals used were of pharmaceutical grade.