Abstract
Recently we reported that the pyridinylimidazole class of p38 mitogen-activated protein (MAP) kinase inhibitors potently inhibited the facilitated transport of nucleosides and nucleoside analogs in K562 cells. These compounds competed with the binding of nitrobenzylthioinosine (NBMPR) to K562 cells, consistent with inhibition of the NBMPR-sensitive equilibrative transporter (ENT1). In this study we examined a large number of additional protein kinase inhibitors for their effects on nucleoside transport. We find that incubation of K562 cells with tyrosine kinase inhibitors (AG825, AG1517, AG1478, STI-571), protein kinase C (PKC) inhibitors (staurosporine, GF 109203X, R0 31-8220, arcyriarubin A), cyclin-dependent kinase inhibitors (roscovitine, olomoucine, indirubin-3′-monoxime), or rapamycin resulted in a dose-dependent reduction of intracellular uptake of [3H]uridine. In contrast, neither the MAP kinase kinase inhibitors (U0126, PD 98059) nor the phosphatidyl inositol-3 kinase inhibitors (wortmannin, LY 294002) affected this process. Furthermore, both transient uptake and prolonged [3H]thymidine incorporation in K562 cells were inhibited by protein kinase inhibitors, inactive analogs of kinase inhibitors (R0 31-6045, SB202474), and NBMPR, independently of effects on cell proliferation as determined by MTT assay. These studies demonstrate that a wide variety of protein kinase inhibitors affect nucleoside uptake through selective inhibition of nucleoside transporters, independently of kinase inhibition.
Footnotes
-
This work was supported by National Institutes of Health Grants R01-GM 59767, AHA EI grant (to L.M.G.), a Leukemia Research Foundation grant (to M.H.), and Grant R01-CA34085 (to B.S.M.).
-
DOI: 10.1124/jpet.102.044214
- Abbreviations:
- PKI
- protein kinase inhibitor
- PKC
- protein kinase C
- dFdC
- gemcitabine
- Ara-C
- cytarabine
- CdA
- cladribine
- F-ara-A
- fludarabine
- CNT
- concentrative nucleoside transporter
- ENT
- equilibrative nucleoside transporter
- NBMPR
- nitrobenzylmercaptopurine ribonucleoside
- MAPK
- mitogen-activated protein kinase
- VEGF
- vascular endothelial growth factor
- DMSO
- dimethyl sulfoxide
- PTK
- protein tyrosine kinase
- TOR
- target-of-rapamycin
- EGFR
- endothelial growth factor receptor
- MTT
- 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium
- SB203580
- 4-(4-fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridinyl)imidazole
- SB203580-iodo
- 4-(3-iodophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)-1H-imidazole
- SB220025
- 5-(2-amino-4-pyrimidinyl)-4-(4-fluorophenyl)-1-(4-piperidinyl)imidazole
- SB202474
- 4-(ethyl)-2-(4-methoxyphenyl)-5-(4-pyridyl)-1H-imidazole
- PD153035 (AG1517)
- 4-(3-bromophenyl)amino-6,7-dimethoxyquinazoline
- AG 1478
- 4-(3-chloroanilino)-6,7-dimethoxyquinazoline
- WHI-P154
- 4-[(3-bromo-4-hydroxyphenyl)amino]-6,7-dimethoxyquinazoline
- WHI-P131
- 4-(4-hydroxyphenyl)amino-6,7-dimethoxyquinazoline
- WHI-P258
- 4-phenylamino-6,7-dimethoxyquinazoline
- WHI-P180
- 4-(3-hydroxyphenyl)amino-6,7-dimethoxyquinazoline
- WHI-P97
- 4-(3,5-dibromo-4-hydroxyphenyl)amino-6,7-dimethoxyquinazoline
- AG18
- α-cyano-(3,4-dihydroxy)cinnamonitrile
- AG490
- α-cyano-(3,4-dihydroxy)-N-benzylcinnamide
- AG 1879 (PP2)
- 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine
- H89 dihydrochloride
- N-[2-((p-bromocinnamyl)amino)ethyl]-5-isoquinolinesulfonamide 2HCl
- ZM336372
- N-[5-(3-dimethylaminobenzamido)-2-methylphenyl]-4-hydroxybenzamide
- KN93
- 2-[N-(2-hydroxyethyl)]-N-(4-methoxybenzenesulfonyl)]amino-N-(4-chlorocinnamyl)-N-methylbenzylamine)
- LY 294002
- 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
- arcyriarubin A
- 2,3-bis(1H-indol-3-yl)maleimide
- Ro 31-8220
- 3-[1-[3-(amidinothio)propyl-1H-indol-3-yl]-3-(1-methyl-1H-indol-3-yl)maleimide
- Ro 32-0432
- 2-{8-[(dimethylamino)methyl]-6,7,8,9-tetrahydropyrido[1,2-α]indol-3-yl}-3-(1-methyl-1H-indol-3-yl)maleimide HCl
- Ro 31-6045
- 2,3-bis(1H-indol-3-yl)-N-methylmaleimide
- GF 109203X
- 2-[1-(3-dimethylaminopropyl)-1H-indol-3-yl]-3-(1H-indol-3-yl)-maleimide
- olomoucine
- 2-(2-hydroxyethylamino)-6-benzylamino-9-methylpurine
- roscovitine
- 2-(R)-(1-ethyl-2-hydroxyethylamino)-6-benzylamino-9-isopropylpurine
- The American Society for Pharmacology and Experimental Therapeutics
JPET articles become freely available 12 months after publication, and remain freely available for 5 years.Non-open access articles that fall outside this five year window are available only to institutional subscribers and current ASPET members, or through the article purchase feature at the bottom of the page.
|
