The percentages of viable (a, d, g), apoptotic (b, e, h) and necr

The percentages of viable (a, d, g), apoptotic (b, e, h) and necrotic Selleck JQEZ5 cells (c, f, i) were determined by FACS-analysis for Annexin V-FITC and Propidiumiodide. Values are means ± SEM of 4 (HT29 and Chang Liver) and 12 (HT1080) independent experiments with consecutive passages. Asterisk symbols on brackets indicate differences between MEK inhibitor treatment groups. *** p ≤ 0.001, ** p ≤ 0.01, * p ≤ 0.05 (one-way ANOVA). Figure 5 Effects of DL-buthionin-(S,R)-sulfoximine on Taurolidine induced cell death in HT29, Chang Liver and HT1080 cells. HT29 (a-c), Chang Liver (d-f) and HT1080 cells (g-i) were incubated with either the glutathione depleting agent DL-buthionin-(S,R)-sulfoximine(BSO)

(1 mM), Taurolidine (TRD) (250 μM) or the combination of both agents (TRD 250 μM + BSO 1 mM) and with Povidon 5% (control) for 24 h. The percentages of viable (a, d, g), apoptotic (b, e, h) and necrotic cells (c, f, i) were determined by FACS-analysis for Annexin V-FITC and Propidiumiodide. Values are

means ± SEM of 9 (HT29 and HT1080) and 4 (Chang Liver) independent experiments with consecutive click here passages. Asterisk symbols on brackets indicate differences between treatment groups. *** p ≤ 0.001, ** p ≤ 0.01, * p ≤ 0.05 (one-way ANOVA). Table 2 Effect of N-Acetylcystein, DL-buthionin-(S,R)-sulfoximine or z-VAD co-incubation with Taurolidine in different cell lines.     HT29 Chang Liver HT1080 AsPC-1 BxPC-3 NAC+TRD 6 h Viable: Ø Ø Ø CoProt Ø   Apo/Nec: Apo⇓ Ø Nec⇓ Nec⇓ Ø NAC+TRD 24 h Viable: CoProt PaProt. Del PaProt PaProt   Apo/Nec: Apo⇓ Apo⇓ Apo⇑ Nec⇑ Nec⇓ Apo⇑ Nec⇓ BSO alone 6 h Viable: Ø Ø Ø Ø Ø   Apo/Nec

Ø Ø Ø Ø Ø BSO+TRD 6 h Viable: Ø Ø Ø Ø Del   Apo/Nec: Ø Ø Nec⇓ Nec⇑ Apo⇓ Nec⇑ BSO alone 24 h Viable: Del Ø Ø Ø Del   Apo/Nec: Nec⇑ Ø Ø Ø Nec⇑ BSO+TRD 24 h Viable: Del MG 132 Ø Ø Del Del   Apo/Nec: Nec⇑ Ø Ø Nec⇑ Apo⇑ Nec⇓ z-VAD+ TRD 24 h Viable: CoProt PaProt PaProt Ø Ø   Apo/Nec: Apo⇓ Ø Nec⇓ Nec⇑ Nec⇓ Effect of N-Acetylcystein (NAC), DL-buthionin-(S,R)-sulfoximine (BSO) or z-VAD co-incubation with Taurolidin (TRD) in different cell lines measured by FACS analysis (Annexin V/Propidium Iodide). NAC = N-Acetylcysteine BSO = DL-buthionin-(S,R)-sulfoximine TRD = Taurolidine Viable = viable cells Apo = apoptotic cells Nec = necrotit cells Ø = no significant effect ⇓ = significant decrease ⇑ = significant increase CoProt. = complete protection PaProt. = partial protection Del. = deleterious In AsPC-1 cells, NAC co-incubation was characterized by a strong reduction of necrosis compared to TRD alone (fig. 6c). Together with a small – but significant – increase in apoptotic cells (fig. 6b) this effect led to a significant increase in viable cells compared to TRD alone (fig. 6a). However, there was no complete recovery in the proportion of viable cells compared to untreated controls (fig. 6a). For that reason the effect could only be designated as partial protection (table 2).

Comments are closed.