Autophagy Pathway Is Required for IL-6 Induced Neuroendocrine Differentiation and Chemoresistance of Prostate Cancer LNCaP Cells
Figure 4
Knockdown of beclin1 suppresses IL-6 induced NED under androgen deprivation conditions.
(A) LNCaP-TR-shBeclin1 cells were treated with 1 µg/ml Dox for 48 hours. TCLs were prepared and immunoblotted to detect beclin1 using GAPDH as the loading control. (B) LNCaP-TR-shBeclin1 cells were treated for 48 hours with 1 µg/ml Dox in order to induce knockdown of beclin1 and they were then treated for another 48 hours with 2.5% CDT or 2.5% CDT plus 100 ng/ml IL-6 to induce cell NED. The inhibition of neurite elongation by beclin1 knockdown was assessed using brightfield microscopy images (40× magnification). (C) The neurite elongation was quantified using the average from 3–5 microscopic fields; bars, SD. (D) TCLs were obtained from LNCaP-TR-shBeclin1 cells treated as described in (A) and these were then analyzed by immunoblotting using the indicated antibodies.