Metabolism and antiproliferative effects of sulforaphane and broccoli sprouts in human intestinal (Caco-2) and hepatic (HepG2) cells
The purpose of this work was to study the absorption and metabolism of sulforaphane (SFN) from broccoli sprouts, their major glucosinolate glucoraphanin and its hydrolysis product SFN. This was done by monitoring SFN’s main metabolites, i.e. SFN, SFN-glutathione and SFN-cysteine, in two different cell models of absorption and metabolism (Caco-2 colorectal carcinoma cells and HepG2 hepatocellular carcinoma cells), during 3, 6, and 24 h of treatment, using a selective UHPLC-QqQ-MS/MS procedure. Concentrations ranging 0.5–90 nmol/l were found within the cells and released in the medium, depending on the type of analyte under study. Cells were capable of conjugative metabolism, since the SFN mercapturic derivatives could be identified in the cell medium. The antiproliferative activity of broccoli sprouts, glucoraphanin and sulforaphane was compared in Caco-2 and HT-29 human colorectal carcinoma cells, and HepG2 hepatocellular carcinoma cells, establishing the minimal concentration of a given compound to achieve half inhibition of the maximal cell growth (IC50) for broccoli sprouts extract and sulforaphane. However, glucoraphanin did not show an antiproliferative effect in the cells under study.
KeywordsBrassica oleraceae Metabolism Cytotoxicity UHPLC-QqQ-MS/MS
This work was supported by the Spanish Ministry of Economy and Competitiveness through projects: AGL2012-30803 and AGL2013-46247-P, and the CYTED Program (Ref. 112RT0460) CORNUCOPIA Thematic Network. N. Baenas was funded by a FPU grant from the Spanish Ministry of Education Fellowship Programme. J. M. Silvan wishes to thank CSIC and FSE for a JAE-Doc postdoctoral contract. The glucoraphanin for these studies was a generous gift from Dr. Gina R. de Nicola and Dr. Renato Iori CRA-CIN, Bologna, Italy.
- Baenas N, Moreno DA, Garcia-Viguera C (2012) Selecting sprouts of brassicaceae for optimum phytochemical composition. J Agric Food Chem 60(45):11409–11420Google Scholar
- Bonnesen C, Eggleston IM, Hayes JD (2001) Dietary indoles and isothiocyanates that are generated from cruciferous vegetables can both stimulate apoptosis and confer protection against DNA damage in human colon cell lines. Cancer Res 61(16):6120–6130Google Scholar
- Cramer JM, Jeffery EH (2011) Sulforaphane absorption and excretion following ingestion of a semi-purified broccoli powder rich in glucoraphanin and broccoli sprouts in healthy men. Nutr Cancer 63(2):196–201Google Scholar
- Kim BR, Hu R, Keum YS, Hebbar V, Shen G, Nair SS, Kong AN (2003) Effects of glutathione on antioxidant response element-mediated gene expression and apoptosis elicited bysulforaphane. Cancer Res 63(21):7520–7525Google Scholar
- Mellon RA, Bennett RN, Holst B, Williamson G (2002) Intact glucosinolate analysis in plant extracts by programmed cone voltage electrospray LC/MS: performance and comparison with LC/MS/MS methods. Anal Biochem 306(1):83–91Google Scholar
- Petri N, Tannergren C, Holst B, Mellon FA, Bao Y, Plumb GW, Bacon J, O’Leary KA, Kroon PA, Knutson L, Forsell P, Eriksson T, Lennernas H, Williamson G (2003) Absorption/metabolism of sulforaphane and quercetin, and regulation of phase II enzymes, in human jejunum in vivo. Drug Metab Dispos 31(6):805–813CrossRefPubMedGoogle Scholar
- Qazi A, Pal J, Mi Maitah, Fulciniti M, Pelluru D, Nanjappa P, Lee S, Batchu RB, Prasad M, Bryant CS, Rajput S, Gryaznov S, Beer DG, Weaver DW, Munshi NC, Goyal RK, Shammas MA (2010) Anticancer activity of a broccoli derivative, sulforaphane, in barrett adenocarcinoma: potential use in chemoprevention and as adjuvant in chemotherapy. Transl Oncol 3(6):389–399PubMedCentralCrossRefPubMedGoogle Scholar
- Singh SV, Herman-Antosiewicz A, Singh AV, Lew KL, Srivastava SK, Kamath R, Brown KD, Zhang L, Baskaran R (2004) Sulforaphane-induced G2/M phase cell cycle arrest involves checkpoint kinase 2-mediated phosphorylation of cell division cycle 25C. J Biol Chem 279(24):25813–25822CrossRefPubMedGoogle Scholar