Effects of aflatoxin B1 on DNA damage and P53geneamplification in hepatocyte-like cells differentiated from mesenchymal stem cells and CD34+ cellsobtained fromhuman umbilical cord blood

Differentiation ofmesenchymal stem cells (MSCs) to hepatocyte-like cells could be associated with development of liver function factors. The impact of differentiation-dependent changes on DNA integrity is not well understood. In this study, hepatocytes and their progenitor stem cells were treated with aflatoxin B1 (AFB1) and amplification of selected genes linked to DNA damage was examined.
MSCs and CD34 cells isolated from umbilical cord blood (UCB) were treated with AFB1 (0, 2.5, 10 and 20 µM) in selective media supporting the hepatocyte differentiation. After 24 htreatment the DNA damage (Comet assay) and amplification rates ofP53 and β-globin genes were measured using real time polymerase chain reaction (QPCR).
The results show that AFB1 treatments resulted in a concentration- dependent increase in the DNA damage and suppression of the specific gene amplification. The extent of DNA damage was significantly greater in hepatocytes differentiated from MSCs when compared to those obtained from CD34 cells. The effects of AFB1 on the rate of selected gene amplification in QPCR showed that the lesions (expressed as lesions/10 kb) in P53 and β-globin genes was significantly greater in hepatocytes derived from MSCs as compared to the cells derived from CD34 cells.
These data together with the results of cytochrome P450 (CYP3A4) expression in the cells suggest that the non-differentiated stem cells are probably less vulnerable to genotoxic agents as compared to hepatocytes differentiated from them.