Research Article

Hsp90 Inhibitor SNX-2112 Enhances TRAIL-Induced Apoptosis of Human Cervical Cancer Cells via the ROS-Mediated JNK-p53-Autophagy-DR5 Pathway

Figure 5

DR5 expression is regulated by SNX-2112-induced autophagy. (a, b) HeLa cells were treated with either TRAIL (200 ng/mL) or SNX-2112 (125 nM) alone or in combination for 48 h. Western blotting was performed to detect the levels of LC3 and Beclin1. Densitometry analyses of the bands for each protein were performed. (c-f) HeLa cells were treated with SNX-2112 for the indicated time (12, 24, and 48 h) or concentration (0, 14, 42, and 125 nM). Western blotting was performed to detect the levels of LC3 and Beclin1. Densitometry analyses of the bands for each protein were performed. (g, h) HeLa cells were treated with SNX-2112 (125 nM) alone or in combination with TRAIL (200 ng/mL) for 48 h. Western blotting was performed to detect the levels of LC3, p-Akt, Akt, p-mTOR, mTOR, p-S6, and p-4EBP1. Densitometry analyses of the bands for each protein were performed. (i, j) HeLa cells were treated with SNX-2112 for the indicated concentration (0, 14, 42, and 125 nM). Western blotting was performed to detect the levels of LC3, p-mTOR, mTOR, p-S6, and p-4EBP1. Densitometry analyses of the bands for each protein were performed. (k, l) HeLa cells were pretreated with BFA (50 nM) for 2 h and then treated with SNX-2112 for 48 h. LC3, DR5, and c-PARP were detected by Western blotting. Densitometry analyses of the bands for each protein were performed. (m, n) HeLa cells were transfected with control siRNA (N-siRNA) or Atg7 siRNAs. After treatment with SNX-2112 (125 nM) for 48 h, Western blotting was used to analyze whole-cell extracts. The protein levels of Atg7, LC3, DR5, and c-PARP were determined using the specific antibody. Densitometry analyses of the bands for each protein were performed. (o, p) The resultant cells were exposed to SNX-2112 (125 nM) in either the absence or the presence of TRAIL (200 ng/mL) for 48 h. Cells were stained with Annexin V/PI, and cell death was measured by FACS. Data are represented as . Error bars represent SD from three separate experiments. and compared with the control group.
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