Carcinogenesis Advance Access originally published online on March 11, 2004
Carcinogenesis 2004 25(8):1427-1433; doi:10.1093/carcin/bgh138
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Carcinogenesis vol.25 no.8 © Oxford University Press 2004; all rights reserved.
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Anthocyanins induce cell cycle perturbations and apoptosis in different human cell lines
Dipartimento di Medicina Sperimentale, sez. Patologia Generale, Universitá di Pavia, and 1 Istituto di Genetica Molecolare C.N.R., I-27100 Pavia, Italy
2 To whom correspondence should be addressed Email: roberto.pizzala{at}unipv.it
To investigate the mechanistic basis for the biological properties of anthocyanins, two aglycone anthocyanins [delphinidin (DY) and cyanidin (CY)] were used to examine their effects on cell cycle progression and on induction of apoptosis in human cancer cells (uterine carcinoma and colon adenocarcinoma cells) and in normal human fibroblasts. These compounds differ in the number and position of hydroxyl groups on the ß ring in the molecular structure. Cellular uptake of anthocyanins was confirmed by HPLC analysis and no metabolites were detected. The clonogenic assay showed that CY induces a dose-dependent growth inhibitory effect only in fibroblasts. This effect was confirmed by flow cytometric analysis, showing a significant reduction of cells in S phase. In contrast, DP inhibited cell growth in normal and tumour cell lines. This event is accompanied in fibroblasts by an accumulation of cells in the S phase suggesting a block in the transition from S to G2 phase. On the other hand, in tumour cell lines we observed a reduction of cells in G1 phase, paralleled by the appearance of a fraction of cells with a hypodiploid DNA content, thus demonstrating an apoptotic effect by DP. The occurrence of apoptosis induced by DP was confirmed by morphological and biochemical features, including nuclear condensation and fragmentation, annexin V staining, DNA laddering and poly(ADP-ribose) polymerase-1-proteolysis. Furthermore, the mitochondrial membrane potential of apoptotic cells after treatment with DP was significantly lost. The different effects exerted by DP as compared with CY suggest that the presence of the three hydroxyl groups on the ß ring in the molecular structure of DP may be important for its greater biological activity.
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