British Journal of Nutrition

Full Papers

Response of cell cycle/stress-related protein expression and DNA damage upon treatment of CaCo2 cells with anthocyanins

Marcella Renisa1, Laura Calandraa1, Christian Scifoa1, Barbara Tomaselloa1, Venera Cardilea2, Luca Vanellaa1, Roberto Beia3, Luca La Faucia4 and Fabio Galvanoa1 c1

a1 Department of Biological Chemistry, Medical Chemistry and Molecular Biology, University of Catania, V.le A. Doria 6, Catania 95125, Italy

a2 Department of Physiological Science, University of Catania, V. le A. Doria 6, Catania 95125, Italy

a3 Department of Experimental Medicine and Biochemical Sciences, University of Rome ‘Tor Vergata’, Via Montpellier 1, I-00133 Rome, Italy

a4 STAFA Department, Mediterranean University of Reggio Calabria, Loc. Feo de Vito, Reggio Calabria 89122, Italy


Anthocyanins are a class of flavonoids, widely spread throughout the plant kingdom, exhibiting important antioxidant and anti-inflammatory actions as well as chemotherapeutic effects; nonetheless, little is known about the molecular mechanisms by which these activities are exerted. The present study is aimed at investigating molecular mechanisms involved in the chemotherapeutic effects induced by both cyanidin-3-O-β glucopyranoside (CY3G) and its aglycon form, cyanidin chloride (CY), in human colon cancer cells (CaCo2). The effect on cell growth, reactive oxygen species (ROS) formation and cell cycle/stress proteins modification, including ataxia teleangectasia mutated protein (ATM), p53, p21, 8-oxoguanine DNA glycosylase (OGG1), 70 kDa heat shock protein (HSP70) and topoisomerase IIβ, as well as on DNA fragmentation, was determined. CY and CY3G treatment affect cell growth and cell proliferation, this latter in a moderately dose-dependent way. Interestingly, ROS level is decreased by any concentration of CY and, only at the lowest concentration, by CY3G. Moreover, the two molecules exert their activities increasing ATM, topoisomerase II, HSP70 and p53 expression. The analysis of DNA fragmentation by Comet assay evidences: (1) a dose-dependent increase in DNA damage only after treatment with CY3G; (2) a more evident trend in the DNA fragmentation when the treatment is performed on agarose embedded cells (cellular atypical Comet); (3) a highly dose-dependent DNA fragmentation induced by CY when the treatment is carried out on agarose embedded naked DNA (acellular atypical Comet). The present findings substantiate a possible chemotherapeutic role of anthocyanins and suggest that CY and CY3G act on CaCo2 by different mechanisms, respectively, ROS-dependent and ROS-independent.

(Received April 18 2007)

(Revised September 28 2007)

(Accepted October 03 2007)

(Online publication December 06 2007)


c1 Corresponding author: Professor Fabio Galvano, fax+39 095 351201, email


Abbreviations: ATM, ataxia teleangectasia mutated protein; CY, cyanidin chloride; CY3G, cyanidin-3-O-β glucopyranoside; DMSO, dimethyl sulphoxide; HSP70, 70 kDa heat shock protein; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; OGG1, 8-oxoguanine DNA glycosylase; ROS, reactive oxygen species; TDNA, percentage of the fragmented DNA; TMOM, tail moment