Overcoming Basal Autophagy, Kangai Injection Enhances Cisplatin Cytotoxicity by Regulating FOXO3a-Dependent Autophagic Cell Death and Apoptosis in Human Lung Adenocarcinoma A549/DDP Cells

Figure 3

A combination of KAI and cisplatin promoted apoptosis in cisplatin-resistant A549/DDP cells. (a) A549/DDP cells were treated with KAI for 2 h; then, cisplatin (10 μM) was added for combined treatment for another 24 h. The effects of cotreatment with KAI and cisplatin on the cell cycle of A549/DDP cells were detected by flow cytometry. (b) The nuclear chromatin condensation of A549/DDP cells was observed by fluorescence microscopy following staining with Hoechst 33258. (c) Annexin V-FITC/PI double staining of A549/DDP cells was performed to detect apoptosis by flow cytometry. (d) Western blot analysis showed apoptosis-related proteins (cleaved caspase-3, p53, Bax, and Bcl-2). β-Actin was used as a loading control. (e) A549/DDP cells were treated with 20 μM of z-Val-Ala-Asp-fluoromethyl ketone for 24 h, then cotreated with KAI and cisplatin. Every experiment result was repeated ≥3 times, and results are presented using values ().

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