RET, the receptor of
glial cell line-derived neurotrophic factor (
GDNF) family ligands, is important for the development of kidney and peripheral neurons.
GDNF promotes survival and differentiation of neurons. Mutation of RET leads to the constitutive signal activation causing
papillary thyroid carcinoma and
multiple endocrine neoplasia type 2 (MEN2). In this study, we report that
GDNF/RET signaling up-regulates
sphingosine kinase (SPHK)
enzyme activity, SPHK1
protein and SPHK1 message in TGW human
neuroblastoma cells. Silencing of SPHK1 using
siRNA inhibited
GDNF-induced neurite formation, GAP43 expression, and cell growth, suggesting the important role of SPHK1 in
GDNF signal transduction. Furthermore, NIH3T3 cells transfected with
MEN2A type mutated RET but not c-RET demonstrated the up-regulation of SPHK activity, SPHK1
protein and SPHK1 message compared with NIH3T3 cells. The cell growth and anchorage-independent colony formation of MEN2A-NIH3T3 was inhibited with
siRNA of SPHK1, while no effect of scramble
siRNA was observed. These results suggest the oncogenic role of SPHK1 in
MEN2A type
tumor. Promoter analysis showed that activator
protein 2 and specificity
protein 1 binding motif of the 5' promoter region of SPHK1 gene is important for its induction by
GDNF. Furthermore, we demonstrated that ERK1/2 and
PI3 kinase are involved in
GDNF-induced SPHK1 transcription by using specific inhibitors.