title: Effect of lentivirus-mediated uPA silencing on the proliferation and apoptosis of chondrocytes and the expression of MMPs.
abstract: The lentivirus-mediated uPA interference in the proliferation, apoptosis, and secretion of osteoarthritic chondrocytes was examined in this study. Cells were obtained from the cartilage tissues of New Zealand white rabbits. They were cultured with interleukin (IL)-1β (10 ng/mL) for 24 h and then divided into three groups: uPA-siRNA group (cells transfected with uPA-siRNA lentiviruses), blank control group (untreated cells), and negative control group (cells transfected with empty vectors). Western blotting and real-time quantitative reverse transcription-PCR (RT-QPCR) were performed to detect the protein and mRNA expression levels of uPA, MMP-1, MMP-3, MMP-9, MMP-10, MMP-13 and MMP-14 in osteoarthritic chondrocytes. Cell Counting Kit-8, flow cytometry, and colony formation assay were used to examine the proliferation and apoptosis of chondrocytes. The results showed that after uPA-siRNA transfection, the protein and mRNA expression levels of uPA, MMP-1, MMP-3, MMP-9, MMP-10, MMP-13, and MMP-14 were significantly decreased (P<0.05 for MMP-1, MMP-9, MMP-10 and MMP-14, P<0.01 for uPA, MMP-3 and MMP-13). Cell proliferation and colony formation rate were significantly higher and the cell apoptosis rate was significantly lower in uPA-siRNA group than in control groups (P<0.01). The proportion of cells in G0/G1 phase was markedly increased and that in the S phase decreased, and the cell cycle was arrested at the G1/S phase in the control group. In the uPA-siRNA group, the proportion of cells in the S phase was significantly increased, resulting in a different proportion of cells in cell cycle phase (P<0.01). It was suggested that the down-regulation of uPA gene could inhibit the expression of MMPs protein and cell apoptosis, increase the proliferation and colony formation of osteoarthritic chondrocytes.
authors: Department of Orthopedics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.|Department of Orthopedics, The First Affiliated Hospital of Medical School, Shihezi University, Shihezi, 832008, China.
authors: Chen-Hui
authors: CH
authors: Shi
authors: Department of Orthopedics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.|Department of Orthopedics, The First Affiliated Hospital of Medical School, Shihezi University, Shihezi, 832008, China.
authors: Wei-Shan
authors: WS
authors: Wang
authors: Department of Orthopedics, The First Affiliated Hospital of Medical School, Shihezi University, Shihezi, 832008, China.|Metabolic Bone Diseases Unit, Division of Endocrinology, Department of Medicine, College of Physicians and Surgeons, Columbia University, New York, 10032, USA.
authors: Zhen-Dong
authors: ZD
authors: Zhang
authors: Department of Orthopedics, The First Affiliated Hospital of Medical School, Shihezi University, Shihezi, 832008, China.|Department of Orthopedic Surgery, Johns Hopkins University School of Medicine, Baltimore, Maryland, 21205, USA.
authors: Chang-Jun
authors: CJ
authors: Li
authors: Department of Orthopedics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
authors: Feng-Jing
authors: FJ
authors: Guo
authors: Department of Orthopedics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
authors: Feng
authors: F
authors: Li
authors: Department of Orthopedics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China. anminchen@tjh.tjmu.edu.cn.
authors: An-Ming
authors: AM
authors: Chen
mesh_terms: D000818:Animals; D017209:Apoptosis; D049109:Cell Proliferation; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology* / Q000201:enzymology; D020868:Gene Silencing; D016086:Lentivirus / Q000235:genetics*; D020782:Matrix Metalloproteinases / Q000378:metabolism*; D011817:Rabbits; D014568:Urokinase-Type Plasminogen Activator / Q000235:genetics*
chemical_list: D014568:Urokinase-Type Plasminogen Activator; D020782:Matrix Metalloproteinases