Figure 7.

Abrogation of BECN1 inhibits autophagy and potentiates paclitaxel cytotoxicity. (A) A2780 and 3AO cells were transfected with BECN1-specific siRNA (line +) or scrambled siRNA (line NC), then were cultured in complete medium, or starved with HBSS (4 h for A2780, 2 h for 3AO), or treated with 10 nM paclitaxel with and without 10 nM BafA1. Whole cell extracts were collected 48 h after siRNA transfection. Protein levels of LC3B and SQSTM1 were determined by western blotting. GAPDH was measured as the loading control. Data were representative of 3 independent experiments. (B) The protein levels of TXNDC17 and BECN1 were detected by western blotting in A2780 and 3AO cells after BECN1 knockdown. (C) A2780 and 3AO cells transfected with BECN1-specific siRNA (line +) or scrambled siRNA (line NC) were seeded in 96-well plates and exposed to paclitaxel at various final concentrations (0, 2, 5, 10, 20, 50 nM; 3 wells for each concentration) for 24 h. Cell viability was measured by CCK-8 kit after 48 h. Data are representative of 3 independent experiments and are expressed as the means ± SD. ** P < 0.01, * P < 0.05.