Phytochemicals play prominent roles in human diet and nutrition as protective redo-active substances in prevention of several disorders and chronic diseases in humans. Today, their function as potent modulators of the mammalian epigenome-regulated gene expression is rapidly emerging. In the present study antiproliferative effects of Indicaxanthin (Ind) from the fruits of Opuntia ficus-indica (1), and potential influence on DNA methylation has been investigated on Caco-2 cells, a human cell line of colorectal carcinoma. Ind caused a clear dose- and time-dependent decrease of the cell proliferation (IC(50) 50 M) associated to apoptosis as demonstrated by phosphatidylserine externalization and depolarization of mithocondrial membrane. Ind decreased the Go-G1phase whereas increased S and G2-M phases of the cell cycle. The phytochemical did not altered the intracellular ROS levels but decreased the [Ca2+]i. Investigation on DNA methylation using MESAP-PCR (Methylation-Sensitive Arbitrarily-Primed Polymerase Chain Reaction) (2), showed that 100 M Ind induced a slight global demethylation after a 48 h treatment. Analysis of epigenetic changes in the DNA methylation pattern at CpG promoter of p16 (INK4a), using MSRE (Methylation-Sensitive Restriction Endonucleases Multiplex-Polymerase Chain Reaction), showed that Ind caused CpG demethylation. Western blotting analysis carried out with p16 monoclonal antibody, confirmed the reactivation of the protein expression. Present data, suggesting that a long-term exposure to indicaxanthin in diet might potentially affect epigenetic machines of the intestinal cells, preventing or repairing initial derangements/disorders, encourage studies on the mechanism involved.
Naselli, F., Modica, M., Attanzio, A., Caradonna, F., Gentile, C., Tesoriere, L., et al. (2012). Pro-apoptotic activity of the phytochemical Indicaxanthin on colorectal carcinoma cells (Caco-2) and epigenetic CpG demethylation of the promoter and reactivation of the expression of p16. In Proceedings of 56th meeting of Italian Society of Biochemistry (pp. 163-163). Milano : Società italiana di Biochimica.
Pro-apoptotic activity of the phytochemical Indicaxanthin on colorectal carcinoma cells (Caco-2) and epigenetic CpG demethylation of the promoter and reactivation of the expression of p16
NASELLI, Flores;ATTANZIO, Alessandro;CARADONNA, Fabio;GENTILE, Carla;TESORIERE, Luisa
;LIVREA, Maria Antonia
2012-01-01
Abstract
Phytochemicals play prominent roles in human diet and nutrition as protective redo-active substances in prevention of several disorders and chronic diseases in humans. Today, their function as potent modulators of the mammalian epigenome-regulated gene expression is rapidly emerging. In the present study antiproliferative effects of Indicaxanthin (Ind) from the fruits of Opuntia ficus-indica (1), and potential influence on DNA methylation has been investigated on Caco-2 cells, a human cell line of colorectal carcinoma. Ind caused a clear dose- and time-dependent decrease of the cell proliferation (IC(50) 50 M) associated to apoptosis as demonstrated by phosphatidylserine externalization and depolarization of mithocondrial membrane. Ind decreased the Go-G1phase whereas increased S and G2-M phases of the cell cycle. The phytochemical did not altered the intracellular ROS levels but decreased the [Ca2+]i. Investigation on DNA methylation using MESAP-PCR (Methylation-Sensitive Arbitrarily-Primed Polymerase Chain Reaction) (2), showed that 100 M Ind induced a slight global demethylation after a 48 h treatment. Analysis of epigenetic changes in the DNA methylation pattern at CpG promoter of p16 (INK4a), using MSRE (Methylation-Sensitive Restriction Endonucleases Multiplex-Polymerase Chain Reaction), showed that Ind caused CpG demethylation. Western blotting analysis carried out with p16 monoclonal antibody, confirmed the reactivation of the protein expression. Present data, suggesting that a long-term exposure to indicaxanthin in diet might potentially affect epigenetic machines of the intestinal cells, preventing or repairing initial derangements/disorders, encourage studies on the mechanism involved.File | Dimensione | Formato | |
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