The ability of hypoxia to modify the gene expression of thymidylate synthase in tumour cells in vivo

被引:10
作者
Ehrnrooth, E
Von der Maase, H
Sorensen, BS
Poulsen, JH
Horsman, MR
机构
[1] Aarhus Univ Hosp, Dept Oncol, DK-8000 Aarhus C, Denmark
[2] Aarhus Univ Hosp, Dept Clin Biochem, DK-8000 Aarhus, Denmark
[3] Aarhus Univ Hosp, Danish Canc Soc, Dept Expt Clin Oncol, DK-8000 Aarhus C, Denmark
关键词
D O I
10.1080/095530099139944
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
Purpose: Hypoxic cells in tumours are resistant to 5-fluorouracil (5-FU). This in vivo study investigated the ability of hypoxia to regulate the gene expression of thymidylate synthase (TS), the target enzyme of 5-FU. Materials and methods: C3H mammary carcinomas, grown in the feet of female CDF1 mice, were used for all experiments. Mice were placed in a 10% oxygen environment for various time periods and the tumour oxygen status was determined with an Eppendorf oxygen electrode. The animals were then injected with BrdU (100 mg/kg, i.p.). Tumours were excised and immediately frozen (-80 degrees C) until isolation of total RNA. The mRNA was reversibly transcribed to complementary DNA and the resulting cDNA amplified in a multiplex PCR reaction, with beta-actin as the internal reference gene. Results: One hour of low oxygen breathing made tumours significantly more hypoxic. This increase was maintained for a maximum incubation period of 48 h. In the same tumours, no change in TS gene expression was seen with up to 3 h of low oxygen breathing. At longer times it decreased, reaching significance at 12-24 h and remaining at this lower level for up to 48 h. BrdU labelling was significantly reduced after breathing low O-2 for 24 h (p = 0.001). Conclusion: Hypoxia-induced down-regulation of TS gene expression was observed. This would be expected to make hypoxic tumour cells more sensitive to 5-FU. Other mechanisms must be responsible for the previously reported resistance to this drug.
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页码:885 / 891
页数:7
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