title: Bortezomib potentially inhibits cellular growth of vascular endothelial cells through suppression of G2/M transition.
abstract: Bortezomib, a selective 26S proteasome inhibitor, has shown clinical benefits against refractory multiple myeloma. The indirect anti-angiogenic activity of bortezomib has been widely recognized; however, the growth-inhibitory mechanism of bortezomib on vascular endothelial cells remains unclear, especially on the cell cycle. Here, we showed that bortezomib (2 nM of the IC(50) value) potently inhibited the cellular growth of human umbilical vascular endothelial cells (HUVECs) via a vascular endothelial growth factor receptor (VEGFR)-independent mechanism resulting in the induction of apoptosis. Bortezomib significantly increased the vascular permeability of HUVECs, whereas a VEGFR-2 tyrosine kinase inhibitor decreased it. Interestingly, a cell cycle analysis using flow cytometry, the immunostaining of phospho-histone H3, and Giemsa staining revealed that bortezomib suppressed the G2/M transition of HUVECs, whereas the mitotic inhibitor paclitaxel induced M-phase accumulation. A further analysis of cell cycle-related proteins revealed that bortezomib increased the expression levels of cyclin B1, the cdc2/cyclin B complex, and the phosphorylation of all T14, Y15, and T161 residues on cdc2. Bortezomib also increased the ubiquitination of cyclin B1 and wee1, but inhibited the kinase activity of the cdc2/cyclin B complex. These protein modifications support the concept that bortezomib suppresses the G2/M transition, rather than causing M-phase arrest. In conclusion, we demonstrated that bortezomib potently inhibits cell growth by suppressing the G2/M transition, modifying G2/M-phase-related cycle regulators, and increasing the vascular permeability of vascular endothelial cells. Our findings reveal a cell cycle-related mode of action and strongly suggest that bortezomib exerts an additional unique vascular disrupting effect as a vascular targeting drug.
authors.affiliation: Department of Genome Biology, Kinki University School of Medicine, Osaka, Japan.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Tamura
authors.forename: Tokuzo
authors.initials: T
authors.lastname: Arao
authors.forename: Kaoru
authors.initials: K
authors.lastname: Tanaka
authors.forename: Hiroyasu
authors.initials: H
authors.lastname: Kaneda
authors.forename: Kazuko
authors.initials: K
authors.lastname: Matsumoto
authors.forename: Kanae
authors.initials: K
authors.lastname: Kudo
authors.forename: Keiichi
authors.initials: K
authors.lastname: Aomatsu
authors.forename: Yoshihiko
authors.initials: Y
authors.lastname: Fujita
authors.forename: Takashi
authors.initials: T
authors.lastname: Watanabe
authors.forename: Nagahiro
authors.initials: N
authors.lastname: Saijo
authors.forename: Yoshikazu
authors.initials: Y
authors.lastname: Kotani
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Nishimura
authors.forename: Kazuto
authors.initials: K
authors.lastname: Nishio
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D001897:Boronic Acids / Q000494:pharmacology*; D000069286:Bortezomib; D016203:CDC2 Protein Kinase; D002199:Capillary Permeability / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D019926:Cyclin B / Q000037:antagonists & inhibitors; D018844:Cyclin-Dependent Kinases; D042783:Endothelial Cells / Q000187:drug effects* / Q000502:physiology; D016195:G2 Phase / Q000187:drug effects*; D006801:Humans; D046988:Proteasome Endopeptidase Complex; D061988:Proteasome Inhibitors; D011719:Pyrazines / Q000494:pharmacology*; D042461:Vascular Endothelial Growth Factor A / Q000502:physiology
chemical_list: D000970:Antineoplastic Agents; D001897:Boronic Acids; D019926:Cyclin B; D061988:Proteasome Inhibitors; D011719:Pyrazines; D042461:Vascular Endothelial Growth Factor A; D000069286:Bortezomib; D016203:CDC2 Protein Kinase; C528899:CDK1 protein, human; D018844:Cyclin-Dependent Kinases; D046988:Proteasome Endopeptidase Complex; C061553:ATP dependent 26S protease