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Effect of Lentivirus-mediated uPA Silencing on the Proliferation and Apoptosis of Chondrocytes and the Expression of MMPs
  • ISSN号:2095-4344
  • 期刊名称:《中国组织工程研究》
  • 时间:0
  • 分类:Q55[生物学—生物化学] Q255[生物学—细胞生物学]
  • 作者机构:[1]Department of Orthopedics, Tongji Hospital Tongji Medical College, Huazhong University of Science and Technology, Wuhan430030, China, [2]Department of Orthopedics, The First Affiliated Hospital of Medical School Shihezi University, Shihezi 832008, China, [3]Metabolic Bone Diseases Unit, Division of Endocrinology, Department of Medicine, College of Physicians and Surgeons, ColumbiaUniversity, New York 10032, USA, [4]Department of Orthopedic Surgery, Johns Hopkins University School of Medicine, Baltimore, Maryland21205, USA
  • 相关基金:supported by grants from the National Natural Science Foundation of China(Nos.81160225,812604531,and 81360451); the Xinjiang Bingtuan Special Program of Medical Science(Nos.2011BC004,2013BA020,and 2014BC003)
中文摘要:

The lentivirus-mediated u PA 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/m L) for 24 h and then divided into three groups: u PA-si RNA group(cells transfected with u PA-si RNA 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 m RNA expression levels of u PA, 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 u PA-si RNA transfection, the protein and mR NA expression levels of uP A, 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 u PA, MMP-3 and MMP-13). Cell proliferation and colony formation rate were significantly higher and the cell apoptosis rate was significantly lower in u PA-si RNA 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 u PAsi RNA 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 uP A gene could inhibit the expression of MMPs protein and cell apoptosis, increase the proliferation and colony formation of osteoarthritic chondrocytes.

英文摘要:

The lentivirus-mediated u PA 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/m L) for 24 h and then divided into three groups: u PA-si RNA group(cells transfected with u PA-si RNA 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 m RNA expression levels of u PA, 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 u PA-si RNA transfection, the protein and mR NA expression levels of uP A, 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 u PA, MMP-3 and MMP-13). Cell proliferation and colony formation rate were significantly higher and the cell apoptosis rate was significantly lower in u PA-si RNA 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 u PAsi RNA 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 uP A gene could inhibit the expression of MMPs protein and cell apoptosis, increase the proliferation and colony formation of osteoarthritic chondrocytes.

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期刊信息
  • 《中国组织工程研究》
  • 北大核心期刊(2014版)
  • 主管单位:中华人民共和国卫生和计划生育委员会
  • 主办单位:中国康复医学会 《中国组织工程研究》杂志社
  • 主编:唐佩福
  • 地址:沈阳浑南新区10002邮政信箱
  • 邮编:110180
  • 邮箱:crter3377@163.com
  • 电话:024-31416864
  • 国际标准刊号:ISSN:2095-4344
  • 国内统一刊号:ISSN:21-1581/R
  • 邮发代号:8-584
  • 获奖情况:
  • 2001“百种中国杰出学术期刊,卫生部首届医药卫生优秀获奖期刊,北方优秀期刊,辽宁省一级期刊,第三、四届沈阳市优秀期刊一等奖
  • 国内外数据库收录:
  • 美国化学文摘(网络版),荷兰文摘与引文数据库,荷兰医学文摘,中国中国科技核心期刊,中国北大核心期刊(2008版),中国北大核心期刊(2014版)
  • 被引量:16688