Regulation of glycolipid sulfotransferase by tyrosine kinases in human renal cancer cells.
Glycolipid sulfotransferase activity in a human renal cancer cell line, SMKT-R3, is enhanced by the action of growth factors such as EGF, TGF-alpha and HGF, whose receptors possess tyrosine kinase domains. We investigated whether tyrosine kinases are involved in the regulation of the sulfotransferase in the cells by using specific tyrosine kinase inhibitors. Genistein and tyrphostin 51 not only cancelled the enhancement of the sulfotransferase by EGF but also reduced the enzyme level to a point much lower than that seen in non-treated cells, whereas they did not affect the sulfotransferase activity in vitro. The activity-reducing effects of genistein were dose- and time-dependent. Genistein also inhibited the cell growth of SMKT-R3 cells. Western blotting using anti-phosphotyrosine monoclonal antibody revealed a tyrosine-phosphorylated protein with an apparent molecular mass of 116 kDa in the non-treated cells. The EGF receptor was tyrosine-phosphorylated by the addition of EGF. The phosphorylations of the 116 kDa protein and EGF receptor were attenuated by co-incubation with genistein. These results indicate that tyrosine kinases including the EGF receptor are involved in the growth of SMKT-R3 cells and in the regulatory mechanisms of glycolipid sulfotransferase in the cells.
Balbaa
M
M
Biochemistry Laboratory, Hokkaido University School of Medicine, Sapporo, Japan.
Honke
K
K
Makita
A
A
D002455:Cell Division / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004815:Epidermal Growth Factor / Q000037:antagonists & inhibitors / Q000494:pharmacology; D066246:ErbB Receptors / Q000037:antagonists & inhibitors; D019833:Genistein; D006801:Humans; D007529:Isoflavones / Q000494:pharmacology; D007680:Kidney Neoplasms / Q000201:enzymology*; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000502:physiology*; D015238:Sulfotransferases / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D004791:Enzyme Inhibitors; D007529:Isoflavones; D004815:Epidermal Growth Factor; D019833:Genistein; D066246:ErbB Receptors; D011505:Protein-Tyrosine Kinases; D015238:Sulfotransferases; C020556:galactosylceramide sulfotransferase