Article Figures & Data
Figures
- Fig. 1.
Expression of TP and DPD in GBM (T) and normal brain (N) tissue. As shown above, TP expression is higher in GBM (n = 9) as compared with normal brain (n = 11) with a mean difference of 6.2 (SD = 7.2; P = 0.13). There was no significant difference in DPD expression levels in the same tissue samples (mean difference of 0.32; SD = 2.13; P = 0.76). The average TP/DPD ratio demonstrated in GBM (2.69) is ∼16-fold higher than that of normal brain tissue (0.17) and is primarily because of TP overexpression in GBM. This profile should result in selective conversion of Capecitabine in tumor compared with normal brain tissues (16-fold higher TP/DPD ratio in GBM relative to normal brain).
- Fig. 2.
Schematic of mouse xenograft location(s) and irradiation. One of the two flanks containing a U87MG glioma xenograft was irradiated, whereas the contralateral flank (containing the other U87MG xenograft) as well as the body of the mouse (including the mouse liver) was lead shielded.
- Fig. 3.
TP expression in irradiated and shielded U87MG glioma xenografts in mice. A, as shown above, TP mRNA levels increased ∼70-fold in irradiated tumors relative to nonirradiated tumors in control mice, remaining elevated 20 days after irradiation. B, of particular interest, TP mRNA levels in the shielded tumors (in the same animal) increased ∼60-fold (relative to initial control levels) 10 and 15 days after irradiation before dropping to control levels at 20 days. Error bars have been incorporated into this figure but are so small as to be obscured by the data point.
- Fig. 4.
TP expression in shielded normal mouse liver. Mouse TP mRNA expression in normal liver was evaluated in shielded and nonirradiated (control) mice. As shown above, there is no significant difference between shielded and nonirradiated mouse liver TP mRNA levels. Error bars have been incorporated into this figure but are so small as to be obscured by the data point.
- Fig. 5.
A, effect of irradiation on mRNA expression of 12 cytokines. Cytokine expression in irradiated tumors (days 0–4 after irradiation) was determined for 12 cytokines using a TaqMan Cytokine Gene Expression Plate I as described in “Materials and Methods.” As shown above, IFN-γ demonstrated the greatest increase in expression (6.3-fold) followed by IL-10 (3.7-fold) in irradiated xenografts relative to control xenografts at 4 days after irradiation. IL-1α demonstrated a <2-fold increase in expression 24 h after irradiation (1.6-fold), returning to baseline levels 2–4 days after irradiation. TNF-α levels (included in the cytokine plate) did not increase above baseline. IL-2, IL-4, IL-5, and IL-8, which are not shown, did not demonstrate any increases in expression after irradiation. IL-1β, IL-12p35, IL-12p40, and IL-15 mRNA levels were beyond the limits of detection in all of the samples (data not shown). B, quantitation of TP in U87MG glioma cells without (−) and with (+) IFN-γ. TP expression (mRNA levels) increased 5-fold in the U87MG glioma cells treated with IFN-γ (+) as compared with untreated cells (−), suggesting a direct effect of IFN-γ on TP transcription.
Tables
- Table 1
Primer and probe sequences and optimal concentrations
Target Sequence Optimal conc. (nm) TP (human) Fwd GGAGAAGGGTGACCGACTCA 100 Probe FAM-CGCTGAGATCAATCGGCCCGACTAT-TAMRA 200 Rev TGCCCAGACTCGGCAAAG 200 TP (mouse) Fwd CGGCCAGAGTGCAAAGCT 200 Probe FAM-CAGCATACAGGATCCCATCAGCAGGAA-TAMRA 200 Rev TCCACAGTGGCTGTCACATCTC 300 TNF-α Fwd GGAGAAGGGTGACCGACTCA 300 Probe FAM-CGCTGAGATCAATCGGCCCGACTAT-TAMRA 250 Rev TGCCCAGACTCGGCAAAG 200 S9 Fwd ATCCGCCAGCGCCATA 100 Probe FAM-AGCAGGTGGTGGTGAACATCCCGTCCTT-TAMRA 300 Rev TCAATGTGCTTCTGGGAATCC 100 DPD Fwd CCAAAGGCAGTAAAGCAGGAA 300 Probe FAM-TGCGCCTGTCACTCTCCATTGCC-TAMRA 25 Rev TCACGACTCCCCGTATCGA 100
Volume 1, Issue 12
