Carcinogenesis Advance Access published online on March 11, 2004
Carcinogenesis, doi:10.1093/carcin/bgh138
© 2004 by Oxford University Press
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MOLECULAR EPIDEMIOLOGY AND CANCER PREVENTION
1 Dipartimento di Medicina Sperimentale, sez. Patologia Generale, 27100 Pavia, Italy
* Corresponding author. E-mail: roberto.pizzala{at}unipv.it.
Received 22 December 2003
; revised 20 February 2004
; accepted 27 February 2004
To investigate the mechanistic basis for the biological properties of anthocyanins, two aglycone anthocyanins (delphinidin and cyanidin) 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
Anthocyanins induce cell cycle perturbations and apoptosis in different human cell lines
2 Istituto di Genetica Molecolare C.N.R., 27100 Pavia, Italy
ring in the molecular structure. Cellular uptake of anthocyanins was confirmed by HPLC analysis and no metabolites were detected. The clonogenic assay showed that cyanidin 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, delphinidin 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 delphinidin. The occurrence of apoptosis induced by delphinidin 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 delphinidin was significantly lost. The different effects exerted by delphinidin as compared to cyanidin suggest that the presence of the three hydroxyl groups on the ring in the molecular structure of delphinidin may be important for its greater biological activity.
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