MLN4924, a novel protein neddylation inhibitor, suppresses proliferation and migration of human urothelial carcinoma: In vitro and in vivo studies
Journal
Cancer Letters
Journal Volume
363
Journal Issue
2
Pages
127-136
Date Issued
2015
Author(s)
Kuo K.-L.
Ho I.-L.
Shi C.-S.
Wu J.-T.
Chou C.-T.
Hsu C.-H.
Hsieh J.-T.
Chang S.-C.
Abstract
MLN4924, a small molecule inhibitor of NEDD8 activating enzyme (NAE), has been reported to elicit an anti-tumor effect on various malignancies. In this study, we investigated the anti-tumor effect of MLN4924 in human urothelial carcinoma (UC) in vitro and in vivo by using three human UC cell lines of various grading (T24, NTUB1 and RT4). The impact of MLN4924 on UC cells was determined by measuring viability (MTT), proliferation (BrdU incorporation), cell cycle progression (flow cytometry with propidium iodide staining) and apoptosis (flow cytometry with annexin V-FITC labeling). The cell cycle regulatory molecules, apoptosis-related molecules, and cell stress-related proteins were examined by Western blotting. The influence of tumor cell migration and invasion was analyzed by Transwell and wound healing assays. We also evaluated the effects of MLN4924 on tumor growth by a SCID xenograft mouse model. The data show that MLN4924 induced dose-dependent cytotoxicity, anti-proliferation, anti-migration, anti-invasion and apoptosis in human UC cells, accompanied by activations of Bad, phospho-histone H2A.X, caspase-3, 7 and PARP, decreased level of phospho-Bcl2, and caused cell cycle retardation at the G2M phase. Moreover, MLN4924 activated endoplasmic reticulum stress-related molecules (caspase-4, phospho-eIF2α, ATF-4 and CHOP) and other stress responses (JNK and c-Jun activations). Finally, we confirmed MLN4924 inhibited tumor growth in a UC xenograft mouse model with minimal general toxicity. We concluded that MLN4924 induces apoptosis and cell cycle arrest, as well as activation of cell stress responses in human UC. These findings imply MLN4924 provides a novel strategy for the treatment of UC. ? 2015 Elsevier Ireland Ltd.
SDGs
Other Subjects
activating transcription factor 4; caspase 3; caspase 4; caspase 7; growth arrest and DNA damage inducible protein 153; histone H2AX; initiation factor 2alpha; lipocortin 5; nicotinamide adenine dinucleotide adenosine diphosphate ribosyltransferase; pevonedistat; propidium iodide; protein c jun; stress activated protein kinase; ((1S,2S,4R)-4-(4-((1S)-2,3-dihydro-1H-inden-1-ylamino)-7H-pyrrolo(2,3-d)pyrimidin-7-yl)-2-hydroxycyclopentyl)methyl sulphamate; cyclopentane derivative; NEDD8 protein, human; pyrimidine derivative; UBA3 protein, human; ubiquitin; ubiquitin protein ligase; aged; animal experiment; animal model; antineoplastic activity; Article; cancer cell destruction; cancer grading; cell cycle progression; cell migration; cell proliferation; cell viability; controlled study; drug mechanism; female; flow cytometry; G2 phase cell cycle checkpoint; human; human cell; in vitro study; in vivo study; M phase cell cycle checkpoint; mouse; nonhuman; priority journal; transitional cell carcinoma; tumor growth; Western blotting; animal; antagonists and inhibitors; apoptosis; carcinoma; cell cycle checkpoint; cell motion; cell proliferation; drug effects; drug screening; genetics; pathology; tumor cell line; Urologic Neoplasms; Animals; Apoptosis; Carcinoma; Cell Cycle Checkpoints; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cyclopentanes; Flow Cytometry; Humans; Mice; Pyrimidines; Ubiquitin-Activating Enzymes; Ubiquitins; Urologic Neoplasms; Xenograft Model Antitumor Assays
Publisher
Elsevier Ireland Ltd
Type
journal article