Fig. 3

ZFP64 reduced the sensitivity of gastric cancer cells to nab-paclitaxel. A. The IC50 value of control and ZFP64-overexpressing AGS cells were measured after treated with nab-paclitaxel for 72 h. B The IC50 value of control and ZFP64-knockdown GC cells after treatment for 72 h. C GC cells were cultured in media and treated with nab-paclitaxel (AGS, 15 nM; MGC-803, 5 nM; HGC-27, 5 nM). After 12, 24, 48 or 72 h of treatment, cell viability was determined by CCK8 assays (n = 6). D GC cells growing in 6-well plates (50,000 cells/well) were exposed to vehicle or nab-paclitaxel (AGS, 15 nM; MGC-803, 5 nM; HGC-27, 5 nM) for 24 h. The cells were trypsinized and collected. One thousand cells from each treatment group were reseeded in 6-well plates. A clonogenic assay was performed. The plates were photographed under a light microscope. E GC cells were grown in duplicate wells of 6-well plates and exposed to vehicle or nab-paclitaxel for 48 h. At the end of the treatment period, cells were trypsinized, and cell apoptosis was evaluated using flow cytometry and an Annexin V/PI Apoptosis Detection Kit. F Schematic representation of the experimental procedure. HGC27 vector cells or HGC27 ZFP64 cells (2 × 10⁶) were subcutaneously injected into athymic nude mice. Fourteen days later, mice were intraperitoneally injected with either vehicle or 10 mg/kg nab-paclitaxel for 3 weeks (twice a week, 8 mice/group). G Representative bioluminescence images of xenografts from mice treated with vehicle or nab-paclitaxel. H-I Tumor growth curves and tumor weights of different treatment groups were analyzed. J TUNEL staining in FFPE samples of tumors from the indicated treatment groups. All experiments in vitro were repeated for 3 times. Data are presented as the means ± SEM. * P < 0.05; ** P < 0.01; *** P < 0.005; **** P < 0.001. FACS, fluorescence-activated cell sorting; TUNEL, terminal-deoxynucleotidyl transferase-mediated nick end labeling; DAPI, 4–6-diamidino-2-phenylindole