Research Article

The Zuo Jin Wan Formula Induces Mitochondrial Apoptosis of Cisplatin-Resistant Gastric Cancer Cells via Cofilin-1

Figure 5

Translocation of cofilin-1 and actin to the mitochondria of SGC7901/DDP cells is induced by ZJW. (a) SGC7901/DDP cells were treated with 50 μg/mL ZJW for 0, 6, 12, and 24 h as indicated. Whole cell lysates and mitochondrial and cytosolic proteins were prepared for the detection of p-cofilin-1, cofilin-1, and gamma-actin using western blotting. The level of each protein was normalised against those of GAPDH (total and cytosolic proteins) or COX IV (mitochondrial proteins). (b) Treated with ZJW (50 μg/mL) for 24 h, a coimmunoprecipitation (co-IP) assay with an anti-cofilin antibody was used to detect the translocation of gamma-actin from the cytoplasm to the mitochondria in SGC7901/DDP cells. (c) Western blot analysis of the G/F-actin ratio changes induced by ZJW in SGC7901/DDP. SGC7901/DDP cells were treated with 50 μg/mL ZJW for 6, 12, and 24 h, respectively, isolating the F-actin extracts and G-actin extracts; the expression of F-actin and G-actin was detected using the anti-F-actin and anti-gamma-actin antibody. or , versus control groups treated ZJW for 0 h. (d) Depolymerisation of F-actin and translocation of G-actin (gamma-actin) from the cytoplasm to the mitochondria was induced by ZJW for 24 h in SGC7901/DDP cells and detected using an immunofluorescence assay. (e) As the same treating method, the ZJW-induced translocation of cofilin-1 from the cytoplasm to the mitochondria (stained with Mitotracker Red) was detected in SGC7901/DDP cells using an immunofluorescence assay.
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