Figure 6

CYP2E1 gene expression is under the transcriptional control of p53. (A) Breast cancer cell lines MCF7, MDA-MB-231, and MDA-MB-468 were treated with ethanol and cellular extracts were submitted to western blot analysis with anti-CYP2E1, anti-p53, and anti-β-actin antibodies. (B) MCF7, MDA-MB-231, T47D and MDA-MB-157 cells were treated with etoposide and total cellular extracts were subjected to western blot analysis as for (A). (C) CYP2E1-Luc reporter assay in MCF7 cells transfected with increasing amounts of CMV-p53. Data presented as fold induction of luciferase activity in cells. Results represent the average of three independent experiments. (D) Chromatin immuniprecipitated using specific p53 monoclonal antibody in non-treated or etoposide-treated MCF7 cells. The pulled-down DNA samples were amplified with PCR primers targeting CYP2E1 promoter. The resultant PCR product was then submitted to 1% agarose gel electrophoresis and visualised by UV light. CYP2E1, cytochrome P450 E1.