TY - JOUR
T1 - UCN-01 alters phosphorylation of Akt and GSK3β and induces apoptosis in six independent human neuroblastoma cell lines
AU - Shankar, Sai Latha
AU - Krupski, Melissa
AU - Parashar, Bhupesh
AU - Okwuaka, Chidi
AU - O'Guin, Kathy
AU - Mani, Sridhar
AU - Shafit-Zagardo, Bridget
PY - 2004/8
Y1 - 2004/8
N2 - In this study we evaluated UCN-01, a small molecule that inhibits protein kinases by interacting with the ATP-binding site, as a potential anti-cancer agent for neuroblastoma. UCN-01 was effective at inducing apoptosis in six neuroblastoma cell lines with diverse cellular and genetic phenotypes. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), terminal deoxynucleotidyl transferase-mediated dUTP nick-end-labeling (TUNEL) assays, detection of active caspase-3 and cleaved poly ADP-ribose polymerase (PARP) confirmed that UCN-01 induced apoptosis. Cell cycle analysis determined that the UCN-01 treated cells accumulated in S phase by 16 h. Unlike vinblastine and docetaxel that increased survivin expression, UCN-01 treatment did not increase X-linked inhibitor of apoptosis protein (XIAP) and survivin levels. Analysis of specific phosphoepitopes on chk1/2, Akt, and GSK3β following UCN-01 treatment determined that there was no significant change in phospho-chk1/2. However, there was decreased immunoreactivity at Ser473 and Thr308 of Akt and Ser9 of GSK3β by 4 h indicating that the Akt survival pathway and downstream signalling was compromised. Thus, UCN-01 was effective at inducing apoptosis in neuroblastoma cell lines.
AB - In this study we evaluated UCN-01, a small molecule that inhibits protein kinases by interacting with the ATP-binding site, as a potential anti-cancer agent for neuroblastoma. UCN-01 was effective at inducing apoptosis in six neuroblastoma cell lines with diverse cellular and genetic phenotypes. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), terminal deoxynucleotidyl transferase-mediated dUTP nick-end-labeling (TUNEL) assays, detection of active caspase-3 and cleaved poly ADP-ribose polymerase (PARP) confirmed that UCN-01 induced apoptosis. Cell cycle analysis determined that the UCN-01 treated cells accumulated in S phase by 16 h. Unlike vinblastine and docetaxel that increased survivin expression, UCN-01 treatment did not increase X-linked inhibitor of apoptosis protein (XIAP) and survivin levels. Analysis of specific phosphoepitopes on chk1/2, Akt, and GSK3β following UCN-01 treatment determined that there was no significant change in phospho-chk1/2. However, there was decreased immunoreactivity at Ser473 and Thr308 of Akt and Ser9 of GSK3β by 4 h indicating that the Akt survival pathway and downstream signalling was compromised. Thus, UCN-01 was effective at inducing apoptosis in neuroblastoma cell lines.
KW - 7-hydroxystaurosporine (UCN-01)
KW - Akt
KW - Apoptosis
KW - GSK3β
KW - Neuroblastomas
KW - Staurosporine
KW - Survivin
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UR - http://www.scopus.com/inward/citedby.url?scp=3342970980&partnerID=8YFLogxK
U2 - 10.1111/j.1471-4159.2004.02543.x
DO - 10.1111/j.1471-4159.2004.02543.x
M3 - Article
C2 - 15255949
AN - SCOPUS:3342970980
SN - 0022-3042
VL - 90
SP - 702
EP - 711
JO - Journal of Neurochemistry
JF - Journal of Neurochemistry
IS - 3
ER -