Bortezomib induces DNA hypomethylation and silenced gene transcription by interfering with Sp1/NF-kappaB-dependent DNA methyltransferase activity in acute myeloid leukemia.
Bortezomib reversibly inhibits 26S proteasomal degradation, interferes with NF-kappaB, and exhibits antitumor activity in human malignancies. Zinc finger protein Sp1 transactivates DNMT1 gene in mice and is functionally regulated through protein abundance, posttranslational modifications (ie, ubiquitination), or interaction with other transcription factors (ie, NF-kappaB). We hypothesize that inhibition of proteasomal degradation and Sp1/NF-kappaB-mediated transactivation may impair aberrant DNA methyltransferase activity. We show here that, in addition to inducing accumulation of polyubiquitinated proteins and abolishment of NF-kappaB activities, bortezomib decreases Sp1 protein levels, disrupts the physical interaction of Sp1/NF-kappaB, and prevents binding of the Sp1/NF-kappaB complex to the DNMT1 gene promoter. Abrogation of Sp1/NF-kappaB complex by bortezomib causes transcriptional repression of DNMT1 gene and down-regulation of DNMT1 protein, which in turn induces global DNA hypomethylation in vitro and in vivo and re-expression of epigenetically silenced genes in human cancer cells. The involvement of Sp1/NF-kappaB in DNMT1 regulation is further demonstrated by the observation that Sp1 knockdown using mithramycin A or shRNA decreases DNMT1 protein levels, which instead are increased by Sp1 or NF-kappaB overexpression. Our results unveil the Sp1/NF-kappaB pathway as a modulator of DNA methyltransferase activity in human cancer and identify bortezomib as a novel epigenetic-targeting drug.
Liu
Shujun
S
Division of Hematology-Oncology, The Ohio State University, Columbus, OH 43210, USA. shujun.liu@osumc.edu
Liu
Zhongfa
Z
Xie
Zhiliang
Z
Pang
Jiuxia
J
Yu
Jianhua
J
Lehmann
Esther
E
Huynh
Lenguyen
L
Vukosavljevic
Tamara
T
Takeki
Mitsui
M
Klisovic
Rebecca B
RB
Baiocchi
Robert A
RA
Blum
William
W
Porcu
Pierluigi
P
Garzon
Ramiro
R
Byrd
John C
JC
Perrotti
Danilo
D
Caligiuri
Michael A
MA
Chan
Kenneth K
KK
Wu
Lai-Chu
LC
Marcucci
Guido
G
D000970:Antineoplastic Agents / Q000494:pharmacology*; D001897:Boronic Acids / Q000494:pharmacology*; D000069286:Bortezomib; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000187:drug effects / Q000378:metabolism*; D019175:DNA Methylation / Q000187:drug effects*; D020868:Gene Silencing / Q000187:drug effects*; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000235:genetics*; D016328:NF-kappa B / Q000502:physiology*; D011494:Protein Kinases / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D011719:Pyrazines / Q000494:pharmacology*; D014158:Transcription, Genetic / Q000187:drug effects*
D000970:Antineoplastic Agents; D001897:Boronic Acids; D016328:NF-kappa B; D011719:Pyrazines; D000069286:Bortezomib; D004248:DNA (Cytosine-5-)-Methyltransferases; D011494:Protein Kinases; C065999:Sp1 kinase