Figure 3

BC sensitizes lung cancer cells to low dose gemcitabine. A , A549 cells infected with BC (filled bars) or control (Con, hatched bars) adenovirus were treated with 0, 0.4, 4, 40, or 400 ng/ml of gemcitabine for 72 h. Cell deaths were determined by subG1 analysis. Open bars represent non-infected A549 cells. The graph was plotted using the means ± SDs of three experiments. *p < 0.001 by the Student's t test for difference in tumor volumes between experimental groups. B and C , A549 and H157 cells infected with BC or control adenovirus were cultured with or without 40 ng/ml gemcitabine for 72 h. Cells were analyzed by flow cytometry for cytochrome C release (B). Apoptotic cell death was determined by FACS analysis of annexin V-PE stained cells among GFP expressing cells. Representative data (upper) and means ± SDs of three experiments (lower) are shown. * p < 0.001 by the Student's t test for differences between amounts of annexin V-PE (+) cells by gemcitabine in control and BC infected cells (C). D , A549 cells infected with BC (filled bars: BC only effect) or Bax (hatched bars) adenovirus were maintained in the presence or absence of 40 ng/ml gemcitabine for 72 h. SubG1 fractions were quantified by PI staining and FACS. Representative data (upper) and the means ± SDs of three experiments (lower) are shown. * p < 0.001 by the Student's t test for differences between amounts of subG1 fraction increases induced by gemcitabine in BC and Bax infected cells. E , A549 cells were transfected with pcDNA, pcDNA-MBC(no tagging GFP), or pcDNA-BAX; 24 hours later, the cells were stained with annexinV-FITC and PI, and cell death was analyzed by flow cytometry. Annexin V-FITC positive cell (without/with-gem), pCDNA: 0.8/1.4(%), MBC: 13.4/20.8(%) and BAX: 22.4/30.6(%). PI positive cell (without/with-gem): pCDNA: 1.7/3.5(%), MBC:13.8/13.7(%) and BAX:11.4/9.4(%). Annexin V-FITC/PI double positive cell (without/with-gem): pCDNA: 0.1/0.3(%), MBC: 11.4/41.7(%) and BAX: 8.3/10.5(%).