Potent cytotoxic effect of a novel nuclear factor-kappaB inhibitor dehydroxymethylepoxyquinomicin on human bladder cancer cells producing various cytokines.
OBJECTIVE
To explore the potential therapeutic effects of the nuclear factor-kappaB (NF-kappaB) inhibitor dehydroxymethylepoxyquinomicin (DHMEQ). KU-19-19 cells, originally derived from a patient with invasive bladder cancer who exhibited marked leukocytosis, produce multiple cytokines. This model of clinically advanced bladder cancer, in which NF-kappaB is constitutively activated, was used in this study.


METHODS
Expression of p65 protein in fractionated KU-19-19 cells was determined by Western blotting analysis. DNA-binding activity of NF-kappaB was detected by electrophoretic mobility shift assay. The cytotoxic effects and induction of apoptosis by DHMEQ were analyzed, and cytokines in the supernatant of KU-19-19 cells cultured with or without DHMEQ were measured by enzyme-linked immunosorbent assay (ELISA). Athymic nude mice bearing KU-19-19 subcutaneous tumors were subjected to intraperitoneal administration of 2 mg/kg/d DHMEQ for 3 weeks. Tumor growth was monitored and microvessel density, vascular endothelial growth factor expression, and the apoptotic index of tumors were evaluated by tissue immunohistochemistry.


RESULTS
NF-kappaB was constitutively activated in KU-19-19 cells. DHMEQ reversibly inhibited the DNA-binding activity of NF-kappaB by blocking its nuclear translocation. Both cell viability and production of cytokines were significantly and dose-dependently suppressed by DHMEQ, and significant apoptosis was also induced. In in vivo studies, the mean tumor volume in mice treated with DHMEQ was significantly smaller than in controls. Immunohistochemical analysis of tumors revealed marked reduction in microvessel density, vascular endothelial growth factor expression, and induction of apoptosis.


CONCLUSIONS
Blockade of NF-kappaB function by DHMEQ may be a useful new molecular targeting treatment for highly aggressive bladder cancer.
Kodaira
Kiichiro
K
Department of Urology, Keio University School of Medicine, Tokyo, Japan.
Kikuchi
Eiji
E
Kosugi
Michio
M
Horiguchi
Yutaka
Y
Matsumoto
Kazuhiro
K
Kanai
Kunimitsu
K
Suzuki
Eriko
E
Miyajima
Akira
A
Nakagawa
Ken
K
Tachibana
Masaaki
M
Umezawa
Kazuo
K
Oya
Mototsugu
M
D000818:Animals; D001549:Benzamides / Q000627:therapeutic use*; D003512:Cyclohexanones / Q000627:therapeutic use*; D016207:Cytokines / Q000096:biosynthesis*; D004195:Disease Models, Animal; D004354:Drug Screening Assays, Antitumor; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D014407:Tumor Cells, Cultured; D001749:Urinary Bladder Neoplasms / Q000188:drug therapy* / Q000378:metabolism*
D001549:Benzamides; D003512:Cyclohexanones; D016207:Cytokines; C464444:dehydroxymethylepoxyquinomicin