Table 1.
Xenograft drug schedule.
Figure 1.
The in vitro effects of DAC and Ara-C in primary paediatric AML.
A). Changes in viability following treatment with DAC or Ara-C at a range of concentrations in 8 primary AML patient samples. B) Changes in proliferation following treatment with DAC or Ara-C at the IC50 dose in 8 primary AML patient cultures. Cells were treated at 0, 24, 48, 72 and 96 h, both cell viability and proliferation were measured at 120 h. All experiments were performed in triplicate.
Table 2.
IC50 dose following treatment with DAC and Ara-C alone.
Figure 2.
Pyrosequencing analysis of 4 tumour suppressor genes following DAC treatment.
Changes in methylation status of a CpG site for 4 representative candidate genes (CSMD1, RARB, CADM1 and CDH13), following treatment with DAC at a range of concentrations in 8 primary AML patient cultures. Cells were treated at 0, 24, 48, 72 and 96 h, DNA methylation was measured at 120 h. All experiments were performed in duplicate.
Figure 3.
The in vitro effects of DAC and Ara-C in combination in primary paediatric AML.
A) The percentage of viable cells following treatment with each drug regimen compared to PBS controls in 5 primary AML patient cultures (* P<0.01). All experiments were performed in triplicate. B) The percentage of cells in each phase of cell cycle following treatment with each drug regimen, for a representative primary AML sample (AML-7). All experiments were performed in duplicate.
Figure 4.
The in vivo effects of DAC and Ara-C in primary paediatric AML xenografts.
A and B) Cells isolated from BM of three xenografts following 5 or 10 days of treatment were stained for human CD45 and CD33 and for mouse CD45; the proportion of human cells which were CD45+CD33+ from each treatment group were compared to Ara-C treatment alone in the BM. C) Cells isolated from the spleen which were human CD45+CD33+ compared to PBS (* P <0.001).
Figure 5.
Q-PCR analysis of candidate genes identified from the expression arrays.
Changes in the expression of 5 genes which were concordantly de-regulated in all three xenografts following sequential treatment with either Ara-C followed by DAC (ILF3 and MARCKS) or DAC followed by Ara-C (FLT3, CTBP1 and MLL5). Assays were carried out in triplicate.
Table 3.
Summary of methylation changes following treatment with different drug regimens.