Figure 2

Inhibition of autophagy enhanced sensitivity of LAD cells to docetaxel. (A, B) LAD cells were treated with 3-methyladenine (3-MA, 5 mM) or transiently transfected with Atg5 or control siRNA. Cells were exposed to the indicated concentrations of docetaxel for 48 h. Viable cells were determined with an MTT assay as described in Materials and methods. (C, D) LAD cells were incubated with various concentrations of docetaxel for 48 h in the presence or absence of 3-MA. The colonies were stained and counted, and survival curves were constructed from three independent experiments. (E) SPC-A1 and H1299 cells were treated with docetaxel (10 μg/l) in the presence or absence of 3-MA (5 mM, 2 h) or Atg5 siRNA. Apoptosis was determined by flow cytometric analysis of Annexin-V/PI staining. (F) SPC-A1/DTX cells were treated with the indicated doses of docetaxel in the presence or absence of 3-MA or Atg5 siRNA. Apoptosis was determined by flow cytometric analysis of Annexin-V/PI staining. Each point or bar represents mean ± S.D. of triplicate determinations. *P< 0.05, **P< 0.01 compared with docetaxel alone treatment.