title: Regulator of cullins-1 expression knockdown suppresses the malignant progression of muscle-invasive transitional cell carcinoma by regulating mTOR/DEPTOR pathway.
abstract: BACKGROUND
  Regulator of cullins-1 (ROC1) is a key subunit in the cullin-RING ligase (CRL) protein complex. Our previous study indicated that ROC1 was essential for bladder cancer cell survival and that ROC1 knockdown inhibited CRL activity, triggering G2 phase arrest and senescence. However, the role of ROC1 in the malignant progression of bladder cancer remained unknown.
  
  
  METHODS
  ROC1 expression in cancer cells was knocked down by siRNA silencing. The effects of ROC1 silencing were evaluated by in vitro assays for cell migration and by an in vivo mouse metastasis model. Epithelial-mesenchymal transition (EMT) induction was evaluated by immunofluorescence staining and western blotting of EMT-associated proteins. ROC1 expression in human tumours was further evaluated by immunohistochemical analysis.
  
  
  RESULTS
  ROC1 knockdown suppresses bladder cancer cell migration by inhibiting EMT. ROC1 knockdown inhibited EMT by inhibiting mammalian target of rapamycin (mTOR) activity via the accumulation of the mTOR-inhibitory protein DEPTOR, a CRL substrate. DEPTOR knockdown partially rescued ROC1 knockdown-inhibited EMT and the ROC1-induced inhibition of cancer cell migration. Furthermore, in vivo studies using a nude mouse metastasis model confirmed the in vitro data. Finally, tissue microarray analysis of clinical bladder cancer specimens indicated a positive correlation between ROC1 expression and EMT.
  
  
  CONCLUSIONS
  ROC1 has an important role in the malignant progression of bladder cancer via the mTOR/DEPTOR pathway. ROC1 may serve as a novel therapeutic target for the treatment of muscle-invasive transitional cell carcinoma.
authors.affiliation: Department of Urology, The Fourth Affiliated Hospital of Nantong University (Yancheng First People's Hospital), Jiangsu 224000, China.
authors.forename: W
authors.initials: W
authors.lastname: Wang
authors.affiliation: Department of Hematology, The Fourth Affiliated Hospital of Nantong University (Yancheng First People's Hospital), Jiangsu 224000, China.
authors.forename: H
authors.initials: H
authors.lastname: Chen
authors.affiliation: Department of Urology, Shanghai First People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200080, China.
authors.forename: Z
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Department of Urology, The Fourth Affiliated Hospital of Nantong University (Yancheng First People's Hospital), Jiangsu 224000, China.
authors.forename: P
authors.initials: P
authors.lastname: Qu
authors.affiliation: Department of Pathology, The Fourth Affiliated Hospital of Nantong University (Yancheng First People's Hospital), Jiangsu 224000, China.
authors.forename: J
authors.initials: J
authors.lastname: Lan
authors.affiliation: Department of Pathology, The Fourth Affiliated Hospital of Nantong University (Yancheng First People's Hospital), Jiangsu 224000, China.
authors.forename: H
authors.initials: H
authors.lastname: Chen
authors.affiliation: Department of Pathology, The Fourth Affiliated Hospital of Nantong University (Yancheng First People's Hospital), Jiangsu 224000, China.
authors.forename: L
authors.initials: L
authors.lastname: Zou
authors.affiliation: Department of Urology, Shanghai First People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200080, China.
authors.forename: J
authors.initials: J
authors.lastname: Qiu
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D015153:Blotting, Western; D002295:Carcinoma, Transitional Cell / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics*; D018450:Disease Progression; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics*; D005260:Female; D005455:Fluorescent Antibody Technique; D055785:Gene Knockdown Techniques; D006801:Humans; D066298:In Vitro Techniques; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D008297:Male; D051379:Mice; D008819:Mice, Nude; D008875:Middle Aged; D009130:Muscle, Smooth / Q000473:pathology; D009361:Neoplasm Invasiveness; D009368:Neoplasm Transplantation; D034741:RNA, Small Interfering; D015398:Signal Transduction; D058570:TOR Serine-Threonine Kinases / Q000378:metabolism*; D046888:Tissue Array Analysis; D001743:Urinary Bladder / Q000473:pathology; D001749:Urinary Bladder Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology
chemical_list: D002352:Carrier Proteins; D047908:Intracellular Signaling Peptides and Proteins; C479529:RBX1 protein, human; D034741:RNA, Small Interfering; C556139:DEPTOR protein, human; C546842:MTOR protein, human; D058570:TOR Serine-Threonine Kinases