Mycotoxin Citrinin Induced Cell Cycle G2/M Arrest and Numerical Chromosomal Aberration Associated with Disruption of Microtubule Formation in Human Cells

Abstract

As a nephrotoxic mycotoxin, citrinin (CTN) contaminates various foodstuffs and animal feed commodities. In the present study, the effects of CTN on cell cycle arrest and microtubule formation were investigated by applying human embryonic kidney (HEK293) cells as a model. Exposure of HEK293 cells to CTN resulted in an arrest of cell cycle G2/M in a concentration-dependent increase. Administrating CTN elevated the expression levels of p53 and p21 proteins, yet attenuated the signals of phosphorylated cell division cycle 2 (cdc2). Furthermore, treating HEK293 with CTN increased both the value of mitotic index and the population of cells recognized by antibody mitotic protein monoclonal 2, suggesting that arrest of CTN-induced cell cycle occurred mainly during the mitotic phase. With the assistance of immunocytostaining of α-tubulin, CTN was found to disrupt the stable microtubule skeleton during the interphase of cell cycle and also interfere with the mitotic spindle integrity during mitosis. Additionally, for either in vivo or in vitro assays, CTN effectively inhibited tubulin polymerization in a concentration-dependent manner. When human peripheral blood mononuclear cells were exposed to CTN, the percentage of cells with numerical chromosome changes was increased by 4.3-fold over that of vehicle-treated group. Results of this study suggest that CTN-activated G2/M arrest primarily arises from the inhibition of tubulin polymerization and associated mitotic spindle formation. Additionally, disruption of microtubule organization by CTN also contributes to the induction of numerical chromosome aberration in human cells.