イトウ　セイジ   ITOU SEIJI   伊藤　誠二    所属   関西医科大学  専門部    職種   名誉教授
論文種別	原著(症例報告除く)
言語種別	英語
査読の有無	査読あり
表題	Controllable urokinase gene expression in trabecular meshwork cells.
掲載誌名	正式名：Journal of clinical and experimental ophthalmology ISSNコード：2155　9570
巻・号・頁	5(2),pp.Article No. 329
著者・共著者	Tsuda　M, Kaneko　S, Ando　A, Nishimura　T, Okuda-Ashitaka　E, Ito　S, Taomoto　M, Matsumura　M, Takahashi　K
発行年月	2014/03
概要	Purpose: Glaucoma is a type of progressive optic neuropathy that finally leads to blindness related to elevated　intraocular pressure (IOP). Accumulation of extracellular matrix in trabecular meshwork (TM) and juxta-canalicular　connective tissues, which form an aqueous outflow pathway, may be a major cause of increased IOP, thus the　fibrinolytic system may be associated with regulation of IOP. We examined the possibility of controllable urokinase　plasminogen activator (uPA) gene transfer into TM cells. Methods: TM cells were freshly isolated from porcine eyes and human TM cells obtained during trabeculectomy procedures and cultured. Total RNA was extracted from human TM cells and reverse transcribed into cDNA. A　reverse-transcribed polymerase chain reaction (RT-PCR) method was then performed to detect the gene expression of uPA. The cDNA of human uPA was sub-cloned into an expression vector (pEYFP-N1 vector) and controllable　expression vector (TRE-Tight vector), then each vector was independently transfected into cultured porcine TM cells. Doxycycline was added to the culture medium to activate the pTet-ON and TRE-Tight vectors. Finally, the　expression of uPA was examined using an enzyme linked immunosorbent assay (ELISA). Results: ELISA findings revealed the expression of uPA (3.5 ng/ml) in medium from cultured porcine TM cells　that had been transfected with the human uPA gene using the pEYFP-N1 vector. Doxycycline induced human uPA　in the pTet-On and TRE-Tight vectors with human uPA gene co-transfected TM cells in a dose-dependent manner. Conclusions: Controllable gene transfer of uPA, which may degrade the extracellular matrix in juxta-canalicular　connective tissue, into TM cells was achieved using a Tet-On system. The present method may be useful as a novel　therapy for glaucoma. DOIコード：10.4172/2155-957-1000329 , 2014