Antiapoptotic function of 17AA(+)WT1 (Wilms' tumor gene) isoforms on the intrinsic apoptosis pathway.
The WT1 gene is overexpressed in human primary leukemia and a wide variety of solid cancers. The WT1 gene is alternatively spliced at two sites, yielding four isoforms: 17AA(+)KTS(+), 17AA(+)KTS(-), 17AA(-)KTS(+), and 17AA(-)KTS(-). Here, we showed that 17AA(+)WT1-specific siRNA induced apoptosis in three WT1-expressing leukemia cell lines (K562, HL-60, and Kasumi-1), but not in WT1-non-expressing lymphoma cell line (Daudi). 17AA(+)WT1-specific siRNA activated caspase-3 and -9 in the intrinsic apoptosis pathway but not caspase-8 in the extrinsic one. On the other hand, 17AA(-)WT1-specific siRNA did not induce apoptosis in the three WT1-expressing cell lines. The apoptosis was associated with activation of proapoptotic Bax, which was activated upstream of the mitochondria. Constitutive expression of 17AA(+)WT1 isoforms inhibited apoptosis of K562 leukemia cells induced by apoptosis-inducing agents, etoposide and doxorubicin, through the protection of mitochondrial membrane damages, and DNA-binding zinc-finger region of 17AA(+)WT1 isoform was essential for the antiapoptotic functions. We further studied the gene(s) whose expression was altered by the expression of 17AA(+)WT1 isoforms and showed that the expression of proapoptotic Bak was decreased by the expression of 17AA(+)KTS(-)WT1 isoform. Taken together, these results indicated that 17AA(+)WT1 isoforms played antiapoptotic roles at some points upstream of the mitochondria in the intrinsic apoptosis pathway.
Ito
K
K
Department of Functional Diagnostic Science, Osaka University Graduate School of Medicine, Suita, Osaka, Japan.
Oji
Y
Y
Tatsumi
N
N
Shimizu
S
S
Kanai
Y
Y
Nakazawa
T
T
Asada
M
M
Jomgeow
T
T
Aoyagi
S
S
Nakano
Y
Y
Tamaki
H
H
Sakaguchi
N
N
Shirakata
T
T
Nishida
S
S
Kawakami
M
M
Tsuboi
A
A
Oka
Y
Y
Tsujimoto
Y
Y
Sugiyama
H
H
D017209:Apoptosis / Q000235:genetics*; D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000502:physiology*; D045744:Cell Line, Tumor; D018922:HL-60 Cells; D006801:Humans; D020014:K562 Cells; D008928:Mitochondria / Q000235:genetics / Q000378:metabolism; D020033:Protein Isoforms / Q000235:genetics / Q000502:physiology; D034741:RNA, Small Interfering / Q000502:physiology; D015398:Signal Transduction / Q000235:genetics*; D025721:WT1 Proteins / Q000235:genetics / Q000502:physiology*
D051017:Apoptosis Regulatory Proteins; D020033:Protein Isoforms; D034741:RNA, Small Interfering; D025721:WT1 Proteins