Antisense inhibition of methylenetetrahydrofolate reductase reduces survival of methionine-dependent tumour lines

被引:12
作者
Sekhon, J
Pereira, P
Sabbaghian, N
Schievella, AR
Rozen, R
机构
[1] McGill Univ, Ctr Hlth, Dept Human Genet, Montreal Childrens Hosp, Montreal, PQ H3Z 2Z3, Canada
[2] McGill Univ, Ctr Hlth, Dept Pediat, Montreal Childrens Hosp, Montreal, PQ H3Z 2Z3, Canada
[3] Vanogen Inc, Cambridge, MA 02139 USA
关键词
methionine-dependence; cancer cells; methylenetetrahydrofolate reductase; antisense technology;
D O I
10.1038/sj.bjc.6600459
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
Transformed cells have been documented to be methionine-dependent, suggesting that inhibition of methionine synthesis might be useful for cancer therapy, Methylenetetrahydrofolate reductase synthesises 5-methyltetrahydrofolate, the methyl donor utilised in methionine synthesis from homocysteine by vitamin B-12-dependent methionine synthase. We hypothesised that methylenetetrahydrofolate reductase inhibition would affect cell viability through decreased methionine synthesis. Using medium lacking methionine, but containing homocysteine and vitamin B, (M-H+), we found that nontransformed human fibroblasts could maintain growth. In contrast, four transformed cell lines (one colon carcinoma, two neuroblastoma and one breast carcinoma) increased proliferation only slightly in the M-H+ medium. To downregulate methylenetetrahydrofolate reductase expression, two phosphorothioate antisense oligonucleotides, EX5 and 677T. were used to target methylenetetrahydrofolate reductase in the colon carcinoma line SW620 400 nM of each antisense oligonucleotide decreased cell survival by approximately 80% (P < 0.01) and 70% (P < 0.0001), respectively, compared to cell survival after the respective control mismatched oligonucleotide. Western blotting and enzyme assays confirmed that methylenetetrahydrofolate reductase expression was decreased. Two neuroblastoma and two breast carcinoma lines also demonstrated decreased survival following EX5 treatment whereas nontransformed human fibroblasts were not affected, This study suggests that methylenetetrahydrofolate reductase may be required for tumour cell survival and that methylenetetrahydrofolate reductase inhibition should be considered for anti-tumour therapy, (C) 2002 Cancer Research UK.
引用
收藏
页码:225 / 230
页数:6
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