Daurisoline Inhibits ESCC by Inducing G1 Cell Cycle Arrest and Activating ER Stress to Trigger Noxa-Dependent Intrinsic and CHOP-DR5-Dependent Extrinsic Apoptosis via p-eIF2α-ATF4 Axis

Figure 5

Daurisoline triggered CHOP-DR5 dependent extrinsic apoptosis. (a) EC1 and ECA109 cells were treated with DAS as described above and were subjected to western blot. (b) The mRNA level of DR5 was quantified by Q-PCR. (c, d) EC1 and ECA109 cells were transfected with control or two siRNA sequences of DR5, then treated with DAS (30 μM) for 24 hours. Apoptosis induction was quantified by Annexin V-FITC/PI double-staining and FACS analysis. DR5 and cleaved PARP protein levels were quantified by western blot. (e, f) EC1 and ECA109 cells were treated with indicated concentrations of DAS for 24 hours, and proteins were extracted and detected by western blot. The mRNA level of CHOP was quantified by Q-PCR. (g, h) EC1 and ECA109 cells were transfected with control or CHOP siRNA, then treated with DAS (30 μM) for 24 hours. Apoptosis induction was quantified by Annexin V-FITC/PI double-staining and FACS analysis. Proteins were extracted and detected by western blot.

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