Figure 1
Ectopic expression of CYP2E1 induces ROS generation in breast cancer cells. (A) MCF7 and MDA-MB-157 cells were treated with ethanol and ROS levels were estimated using H2DCFDA and FACS analysis. (B) MCF7, MDA-MB-231 and MDA-MB-157 cells were treated with etoposide and NAC and ROS levels were estimated as in (A). (C) MCF7, MDA-MB-231 and MDA-MB-157 cells were co-transfected with CD20 and either PCDNA3 or CMV-CYP2E1 expression plasmid. Subsequent to transfection, cells were treated with H2DCFDA fluorescent stain, and transfected cells were sorted with an APC-conjugated anti-CD20 antibody and FACS analysis. Data represent fold induction of ROS levels. Error bars indicate standard deviation obtained from three independent experiments performed in duplicates. Asterisks indicate significant difference at P <0.05. CYP2E1, cytochrome P450 E1; FACS, fluorescence-activated cell sorting; H2DCFDA, 2 2.9.2 2′,7′-dichlorodihydrofluorescein diacetate; NAC, N-acetylcysteine; ROS, reactive oxygen species.