The use of a fluorescent methotrexate probe to monitor the effects of three vinca alkaloids on a mixed population of parental L1210 and gene-amplified methotrexate-resistant cells by flow cytometry
✍ Scribed by David G. Poppitt; Alan T. McGown; Brian W. Fox
- Publisher
- Springer
- Year
- 1984
- Tongue
- English
- Weight
- 374 KB
- Volume
- 13
- Category
- Article
- ISSN
- 0344-5704
No coin nor oath required. For personal study only.
✦ Synopsis
Cells res&tant to methotrexate (LI210/R7A) and possessing an increased level of dihydrofolate reductase due to gene amplification can be detected by the technique of flow cytofluorimetry using a new fluorescent derivative of methotrexate (F-MTX) based on a putrescine linker. Comparative studies of dihydrofolate reductase enzyme and cell growth inhibition following treatment with methotrexate and F-MTX suggest that the two agents possess similar modes of action.
In an artificially mixed population of cells sensitive and resistant to methotrexate it is possible, using F-MTX, to recognise and separate distinct cell subpopulations showing differential fluorescence using a fluorescence-activated cell sorter (FACS IV). The selective removal of the resistant cells within a mixed population of sensitive and resistant cells has been demonstrated for 5 x 10 -8 M vinblastine by means of flow cytometry. The effectiveness of the vinca alkaloids decreases in the order vinblastine > vindesine > vincristine, which previously was shown to be the order of effectiveness in producing collateral sensitivity.