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[Cancer Research 60, 1426-1433, March 1, 2000]
© 2000 American Association for Cancer Research

Tumor Biology

Sulforaphane, a Naturally Occurring Isothiocyanate, Induces Cell Cycle Arrest and Apoptosis in HT29 Human Colon Cancer Cells

Laurence Gamet-Payrastre1, Pengfei Li, Solange Lumeau, Georges Cassar, Marie-Ange Dupont, Sylvie Chevolleau, Nicole Gasc, Jacques Tulliez and François Tercé

INRA, Laboratoire des Xénobiotiques, 31931 Toulouse Cedex, France [L. G-P., P. L., S. L., S. C., N. G., J. T.]; INSERM U326, 31059 Toulouse Cedex, France [F. T.]; INSERM U395, Service Commun d’Analyse et de Tri Cellulaire, 31024, Toulouse Cedex, France [G. C.]; and Laboratoire de Biologie Moléculaire des Eucaryotes, 31062, Toulouse Cedex, France [A. D., M-A. D.]

Sulforaphane is an isothiocyanate that is present naturally in widely consumed vegetables and has a particularly high concentration in broccoli. This compound has been shown to block the formation of tumors initiated by chemicals in the rat. Although sulforaphane has been proposed to modulate the metabolism of carcinogens, its mechanism of action remains poorly understood. We have previously demonstrated that sulforaphane inhibits the reinitiation of growth and decreases the cellular viability of quiescent human colon carcinoma cells (HT29). Moreover, the weak effect observed on differentiated CaCo2 cells suggests a specific anticancer activity for this compound.

Here we investigated the effect of sulforaphane on the growth and viability of HT29 cells during their exponentially growing phase. We observed that sulforaphane induced a cell cycle arrest in a dose-dependent manner, followed by cell death. This sulforaphane-induced cell cycle arrest was correlated with an increased expression of cyclins A and B1. Moreover, we clearly demonstrated that sulforaphane induced cell death via an apoptotic process. Indeed, a large proportion of treated cells display the following: (a) translocation of phosphatidylserine from the inner layer to the outer layer of the plasma membrane; (b) typical chromatin condensation; and (c) ultrastructural modifications related to apoptotic cell death. We also showed that the expression of p53 was not changed in sulforaphane-treated cells. In contrast, whereas bcl-2 was not detected, we observed increased expression of the proapoptotic protein bax, the release of cytochrome c from the mitochondria to the cytosol, and the proteolytic cleavage of poly(ADP-ribose) polymerase. In conclusion, our results strongly suggest that in addition to the activation of detoxifying enzymes, induction of apoptosis is also involved in the sulforaphane-associated chemoprevention of cancer.




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