Focal adhesion kinase-dependent apoptosis of melanoma induced by tyrosine and phenylalanine deficiency.
We found previously that restriction of tyrosine (Tyr) and phenylalanine (Phe) inhibited growth and metastasis of B16BL6 murine melanoma and arrested these cells in the G0-G1 phase of the cell cycle. Here, we report that deprivation of these two amino acids in vitro induces apoptosis in B16BL6 and in human A375 melanoma cells but not in nontransformed, neonatal murine epidermal cells or human infant foreskin fibroblasts. Four days after deprivation of Tyr and Phe in vitro, 37% of B16BL6 and 51% of A375 melanoma cells were undergoing apoptosis. Apoptosis was not associated with elevation in intracellular calcium or alteration in p53 or c-myc protein expression. Expression and Tyr phosphorylation of focal adhesion kinase (FAK) were inhibited in both melanoma cell lines by deprivation of Tyr and Phe but not by deprivation of glutamine or serum. Tyr phosphorylation of FAK in Tyr- and Phe-deprived melanoma cells was enhanced within 30 min of refeeding with complete DMEM. FAK protein expression recovered within 60 min, and cell viability recovered within 24 h. Genistein, a tyrosine kinase inhibitor that specifically inhibits Tyr phosphorylation of FAK, did not induce apoptosis in A375 melanoma cells at a concentration of 50 microM. Genistein prevented the recovery of cell viability upon refeeding with Tyr and Phe to previously deprived A375 melanoma cells. These data collectively indicate that apoptosis induced by Tyr and Phe deprivation is FAK-dependent.
Fu
Y M
YM
Department of Pharmaceutical Sciences, College of Pharmacy, and the Cancer Prevention and Research Center, Washington State University, Pullman 99164-6510, USA.
Yu
Z X
ZX
Pelayo
B A
BA
Ferrans
V J
VJ
Meadows
G G
GG
D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D002118:Calcium / Q000378:metabolism; D015815:Cell Adhesion Molecules / Q000502:physiology*; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; D016193:G1 Phase / Q000502:physiology; D006801:Humans; D007424:Intracellular Fluid / Q000378:metabolism; D008545:Melanoma / Q000378:metabolism / Q000473:pathology*; D008546:Melanoma, Experimental / Q000378:metabolism / Q000473:pathology*; D051379:Mice; D010649:Phenylalanine / Q000172:deficiency*; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000502:physiology*; D016271:Proto-Oncogene Proteins c-myc / Q000096:biosynthesis; D016192:Resting Phase, Cell Cycle / Q000502:physiology; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis; D014443:Tyrosine / Q000172:deficiency*