Figure 2
From: ZnCl2 sustains the adriamycin-induced cell death inhibited by high glucose
The ADR-induced cell death is reduced by HG and re-established by ZnCl2 cotreatment. (a) RKO and HCT116 cells (2 × 105) were plated at subconfluence in culture media containing 10% FBS and 1 g/l D-glucose. The day after, medium was changed with medium containing 2% FBS with either 1 g/l D-glucose (low glucose) or 4.5 g/l D-glucose (HG) for 24 h before adding chemotherapeutic drugs ADR (2 μg/ml) with or without ZnCl2 (100 μM). After 24 h, the percentage of dead cells was scored by Trypan blue exclusion. Data are presented as mean±S.E.M (n=6) (one-way ANOVA plus Bonferroni test, **P<0.001 ADR versus Mock and ADR/HG/ZnCl2 versus ADR/HG, *P<0.01 ADR/HG versus ADR). (b) Live images of cells analyzed in (a) were taken before lysing cells. Black arrows indicate dead cells. (c) RKO cells were treated with ADR (2 μg/ml) in low and high glucose condition with or without ZnCl2 (100 μM) for 24 h; the irreversible caspase inhibitor z-VADfmk was used at 40 mM for 16 h. After treatments, cells were in part fixed and stained with PI for sub-G1 evaluation (upper panel) or lysed and analyzed by western immunoblotting to assess PARP cleavage (lower panel) and relative quantification of PARP cleavage/β-actin ratio (right panel). Anti-β-actin was used as protein loading control. The predicted molecular weight is indicated (kDa). For sub-G1 analysis, data are presented as mean±S.D. In the right panel data are presented as mean±S.E.M. (n=6) (one-way ANOVA plus Bonferroni test, *P<0.001 ADR versus Mock, ADR/HG versus ADR, ADR/HG/ZnCl2 versus ADR/HG, ADR+z-VAD versus ADR)
