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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2010.1
- 수록면
- 163 - 171 (9page)
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Purpose The aim of this study is to investigate the effect of genetic variations and the expression of the reduced folate carrier (RFC) and dihydrofolate reductase (DHFR) on the drug sensitivity to methotrexate (MTX) in different cancer cell lines.
Materials and Methods We examined the six human cancer cell lines (MCF-7, AGS, A549, NCI-H23, HCT-116 and Saos- 2). The cytotoxicity of MTX was measured by sulforhodamine B (SRB) assay. The expressions of the DHFR and RFC were evaluated by real-time PCR and western blotting. Four single nucleotide polymorphisms (SNPs) of the DHFR and two SNPs of the RFC were genotyped.
Results The IC50s of MTX was in an extensively broad range from 6.05±0.81 nM to＞1,000 nM in the cell lines. The Saos-2 (＞1,000 nM) and MCF-7 (114.31±5.34 nM) cells were most resistant to MTX; in contrast, the AGS and HCT-116 cells were highly sensitive to MTX with an IC50 of 6.05±0.81 nM and 13.56±3.76 nM, respectively. A reciprocal change of the RFC and DHFR mRNA expression was found between the MTX-sensitive AGS and MTX-resistant Saos-2 cells. There was no significant difference in the expression levels of RFC protein in both the AGS and Saos-2 cells, whereas DHFR protein was more increased in the MTX-resistant Saos-2 cells treated with MTX. The genotype of the MTX-sensitive AGS cells were mutant variants of the DHFR; in contrast, the Saos-2 cells had the wild-type of the DHFR.
Conclusion In conclusion, this study showed that inverse change of the RFC and DHFR mRNA and protein expression was associated with RFC and DHFR polymorphisms and it is postulated that this phenomenon might play an important role in sensitivity of certain cancers to MTX.
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참고문헌 (20)
참고문헌 신청
Affleck JG / 2010 / Selection for methotrexate resistance in mammalian cells bearing a Drosophila dihydrofolate reductase transgene: methotrexate resistance in transgenic mam
Backus HH / 2000 / Folate depletion increases sensitivity of solid tumor cell lines to 5-fluorouracil and antifolates / Int J Cancer 87 : 771 ~ 778
Dulucq S / 2008 / DNA variants in the dihydrofolate reductase gene and outcome in childhood ALL / Blood 111 : 3692 ~ 3700
Gomes CM / 2006 / Multidrug resistance mediated by ABC transporters in osteosarcoma cell lines: mRNA analysis and functional radiotracer studies / Nucl Med Biol 33 : 831 ~ 8
Goto Y / 2001 / A novel single-nucleotide polymorphism in the 3’-untranslated region of the human dihydrofolate reductase gene with enhanced expression / Clin Cancer Res 7 :
Backus HH / 2000 / Folate depletion increases sensitivity of solid tumor cell lines to 5-fluorouracil and antifolates / Int J Cancer 87 : 771 ~ 778
Dulucq S / 2008 / DNA variants in the dihydrofolate reductase gene and outcome in childhood ALL / Blood 111 : 3692 ~ 3700
Gomes CM / 2006 / Multidrug resistance mediated by ABC transporters in osteosarcoma cell lines: mRNA analysis and functional radiotracer studies / Nucl Med Biol 33 : 831 ~ 8
Goto Y / 2001 / A novel single-nucleotide polymorphism in the 3’-untranslated region of the human dihydrofolate reductase gene with enhanced expression / Clin Cancer Res 7 :
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