title: The suppressive effect of Rho kinase inhibitor, Y-27632, on oncogenic Ras/RhoA induced invasion/migration of human bladder cancer TSGH cells.
abstract: Urothelial cell carcinoma is the most common type of malignancy found in long-term dialysis patients and kidney transplant recipients in Taiwan. Surgical specimens of tumorous and non-tumorous bladder tissues were collected from 12 patients with bladder cancer. Increased expressions of Ras, RhoA, Akt, PI-3K were demonstrated in the tumors as compared to adjacent control tissues. To understand the impact of Ras over-expression on bladder cancer progression, human bladder cancer TSGH 8301 cells were transfected with Ras DNA. The Ras-transfected cells were then treated with either a PI-3K inhibitor (wortmannin) or Rho kinase inhibitor (Y-27632) and the expressions of Ras, PI-3K, Akt, NF-kappaB, and RhoA were analyzed. Fluorescent phalloidin staining demonstrated more intense F-actin staining in the Ras over-expressed cells than in the control cells, and the intensity of F-actin was inhibited by Y-27632. A gelatin zymography study demonstrated that the MMP-2 and MMP-9 expressions of the Ras-transfected cells were enhanced, and Y-27632 treatment reduced the levels of MMP-2 and MMP-9. Similarly, a wound healing assay revealed that the ability of cell migration was markedly increased by Ras transfection and the healing rate after treatment of Y-27632 was delayed. Our results provide evidence that Ras-induced RhoA and NF-kappaB activation was involved in the invasion/migration of bladder cancer. Through Ras and/or RhoA inhibition, there might be an opportunity for new therapeutic interventions in bladder cancer.
authors.affiliation: Institute of Medicine, Chung Shan Medical University, Taichung 402, Taiwan; Department of Nephrology, Chung Shan Medical University Hospital, Taichung 402, Taiwan.
authors.forename: Horng-Rong
authors.initials: HR
authors.lastname: Chang
authors.forename: Hui-Pei
authors.initials: HP
authors.lastname: Huang
authors.forename: Yu-Lin
authors.initials: YL
authors.lastname: Kao
authors.forename: Sung-Lang
authors.initials: SL
authors.lastname: Chen
authors.forename: Sheng-Wen
authors.initials: SW
authors.lastname: Wu
authors.forename: Tung-Wei
authors.initials: TW
authors.lastname: Hung
authors.forename: Jong-Da
authors.initials: JD
authors.lastname: Lian
authors.forename: Chau-Jong
authors.initials: CJ
authors.lastname: Wang
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D000577:Amides / Q000494:pharmacology*; D045744:Cell Line, Tumor; D002465:Cell Movement; D003599:Cytoskeleton; D005260:Female; D006801:Humans; D008297:Male; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D008875:Middle Aged; D016328:NF-kappa B / Q000378:metabolism; D009361:Neoplasm Invasiveness; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D011725:Pyridines / Q000494:pharmacology*; D014162:Transfection; D001749:Urinary Bladder Neoplasms / Q000378:metabolism*; D018631:ras Proteins / Q000235:genetics / Q000378:metabolism*; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors*; D020742:rhoA GTP-Binding Protein / Q000037:antagonists & inhibitors / Q000378:metabolism*
chemical_list: D000577:Amides; D016328:NF-kappa B; D047428:Protein Kinase Inhibitors; D011725:Pyridines; C108830:Y 27632; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt; D054460:rho-Associated Kinases; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D018631:ras Proteins; D020742:rhoA GTP-Binding Protein