Activation of FKHRL1 plays an important role in protecting erythroid cells from erythropoietin deprivation-induced apoptosis in a human erythropoietin-dependent leukemia cell line, UT-7/EPO.
FKHRL1 is one of the human homologues of DAF-16, which is concerned with longevity in Caenorhabditis elegans. Previously, we demonstrated that FKHRL1 functions downstream of Akt in erythropoietin (EPO) signaling and that it is directly phosphorylated by activated Akt. Because phosphorylated FKHRL1 loses its transcriptional activity and translocates into the cytoplasm, FKHRL1 appears to be nonfunctional in the presence of EPO. Conversely, EPO deprivation leads to FKHRL1 dephosphorylation and its translocation into the nucleus, suggesting that FKHRL1 becomes active as a transcription factor in the absence of EPO. On the basis of these findings, we hypothesized, by analogy with C elegans, that erythroid cells possess self-defense machinery against life-threatening surroundings. We prepared a dominant-negative mutant of FKHRL1 (FKHRL1-DN) lacking the transactivation domain and prepared FKHRL1 small interfering RNA (siRNA), and we used constructs to transfect a human EPO-dependent cell line, UT-7/EPO. In the parental cells, 24-hour EPO deprivation induced transient cell cycle arrest without apoptosis. On the other hand, stable transfectants expressing FKHRL1-DN or FKHRL1 siRNA underwent rapid apoptosis after EPO deprivation in the UT-7/EPO cells. In conclusion, FKHRL1 activation plays an important role in the extension of survival of erythroid cells after EPO deprivation. This phenomenon appears to correspond to dauer formation in C elegans. Thus, the mechanism of lifespan extension may be broadly conserved from C elegans to humans.
Uchida
Mie
M
Division of Hematology, Department of Medicine, Jichi Medical University, Tochigi, Japan.
Kirito
Keita
K
Endo
Hitoshi
H
Ozawa
Keiya
K
Komatsu
Norio
N
D017209:Apoptosis / Q000187:drug effects*; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D041905:Erythroid Cells / Q000378:metabolism* / Q000473:pathology*; D004921:Erythropoietin / Q000494:pharmacology*; D000071316:Forkhead Box Protein O3; D051858:Forkhead Transcription Factors / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007938:Leukemia / Q000378:metabolism* / Q000473:pathology*; D021381:Protein Transport; D034741:RNA, Small Interfering / Q000235:genetics
C499517:FOXO3 protein, human; D000071316:Forkhead Box Protein O3; D051858:Forkhead Transcription Factors; D034741:RNA, Small Interfering; D004921:Erythropoietin