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Table 3 HPLC analysis of troxacitabine metabolites after exposure to gemcitabine.

From: Synergistic activity of troxacitabine (Troxatyl™) and gemcitabine in pancreatic cancer

Treatment

Troxacitabine incorporation

into DNA

(CPM/106 cells)

Metabolites

(pmol/mg protein)

  

MP

DP

TP

[3H]Troxacitabine 10 μM

[3H]Troxacitabine 10 μM + Gemcitabine 10 μM

1248 ± 153

1226 ± 91

5.0 ± 1.6

2.8 ± 0.9

26.0 ± 7.0

13.0 ± 4.0

6.4 ± 1.5

3.2 ± 1.1

[3H]Troxacitabine 10 μM

[3H]Troxacitabine 10 μM + Gemcitabine 0.1 μM

1248 ± 153

1097 ± 298

5.6 ± 1.5

1.7 ± 2.0

22.0 ± 5.0

23.0 ± 2.0

4.5 ± 1.7

5.2 ± 1.0

[3H]Troxacitabine 300 nM

[3H]Troxacitabine 300 nM + Gemcitabine 0.5 nM

4039 ± 118

3838 ± 509

3.0 ± 0.1

2.0 ± 0.2

6.0 ± 0.2

5.9 ± 0.2

2.6 ± 0.3

2.5 ± 0.2

  1. AsPc-1 cells were exposed to [3H]troxacitabine in absence or presence of gemcitabine as described in Materials and Methods. Cells were harvested by trypsinization, washed in cold PBS and extracted with 10% TCA for 10 min on ice. The acid-insoluble pellets, containing DNA into which [3H]troxacitabine was incorporated, were solubilized in 0.5 N NaOH and their radioactive content was determined. Values are presented as (CPM/106 cells ± SD). The supernatants, containing phosphorylated metabolites of [3H]troxacitabine, were analyzed by HPLC. Values (pmol/mg protein ± SEM) for troxacitabine monophosphate (MP), diphosphate (DP) and triphosphate (TP) are given below.
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BMC Cancer

ISSN: 1471-2407

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