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Biochemical, Molecular, and Genetic Mechanisms

Polyamine Metabolism and Transforming Growth Factor-β Signaling Are Affected in Caco-2 Cells by Differentially Cooked Broccoli Extracts^1–5,

⁶ Institute of Food Research, Norwich Research Park, Colney Lane, Norwich NR4 7UA, UK and ⁷ Centro de Investigação e de Tecnologias Agro-Ambientais e Biológicas-Departamento de Fitotecnia e Engenharia Rural, Universidade de Trás-os-Montes e Alto Douro, 5001-801 Vila Real, Portugal

^* To whom correspondence should be addressed. E-mail: caroline.furniss{at}bbsrc.ac.uk.

The health benefits of consuming cruciferous vegetables are widely considered to be due to the biological activity of glucosinolate degradation products. However, it is conceivable that other phytochemicals within crucifers may also have biological activity that may contribute to health benefits. In this study, we analyzed global gene expression in Caco-2 cells exposed to extracts derived from broccoli that had been heat treated to different extents to result in contrasting profiles of glucosinolates and their degradation products. Extracts microwaved for 0, 1, and 4 min contained 9.5, 25.5, and 0 µmol/L sulforaphane and induced changes in expression of 381, 1017, and 101 genes, respectively (>2 fold; P < 0.01). Seventy-two genes showed similar changes in expression after treatment with all 3 extracts. These included genes involved in polyamine catabolism and transforming growth factor (TGF)-β signaling. Consistent with these changes in gene expression, subsequent studies demonstrated that exposing cells to these extracts, including the 4-min extract that contained no glucosinolate degradation products, increased putrescine and N-acetyl-spermine concentration, and suppressed the TGFβ1-mediated induction of phosphorylated Smad 2. This is the first report, to our knowledge, of phytochemicals from a cruciferous vegetable affecting both a signaling pathway and a catabolic process.