Matrix vesicle plasma cell membrane glycoprotein-1 regulates mineralization by murine osteoblastic MC3T3 cells
Matrix vesicle plasma cell membrane glycoprotein-1 regulates mineralization by murine osteoblastic MC3T3 cells
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DOI：
10.1359/jbmr.1999.14.6.883
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发表时间：
1999-06-01
影响因子：
6.2
通讯作者：
Terkeltaub, R
中科院分区：
文献类型：
--
作者：
Johnson, K;Moffa, A;Terkeltaub, R
产生无机焦磷酸盐（PPi）的核苷三磷酸焦磷酸水解酶（NTPPPH） PC-1的自然无意义截断突变与“踮脚走路”（ttw）小鼠的脊柱和关节周围韧带肥大和软骨钙化有关。因此，我们验证了PC-1直接作用于细胞外基质抑制矿化的假设，培养的成骨细胞MC3T3细胞在12-14天表达PC-1 mRNA并产生羟基磷灰石沉积物。在14 d内，NTPPPH活性稳步增加。转化生长因子- β和1,25-二羟基维生素D-3增加基质囊泡（MVs）中PC-1和NTPPPH，由于PC-1/NTPPPH在矿化MC3T3细胞中受到调节，我们用重组腺病毒稳定转染或感染细胞，使PC-1的表达量增加2至6倍。此外，与转染空质粒或酶失活PC-1的细胞相比，转染PC-1的MC3T3细胞沉积的羟基磷灰石（按重量计）减少了80-90%，来自过表达活性PC-1的细胞的mv中atp依赖性Ca-45的沉淀相对减少。因此，PC-1的调节控制成骨细胞来源的MVs和基质羟基磷灰石沉积的PPi含量和功能。PC-1可能为某些骨矿化疾病提供新的治疗靶点。
A naturally occurring nonsense truncation mutation of the inorganic pyrophosphate (PPi)-generating nucleoside triphosphate pyrophosphohydrolase (NTPPPH) PC-1 is associated with spinal and periarticular ligament hyperostosis and cartilage calcification in "tiptoe walking" (ttw) mice. Thus, we tested the hypothesis that PC-1 acts directly in the extracellular matrix to restrain mineralization, Cultured osteoblastic MC3T3 cells expressed PC-1 mRNA and produced hydroxyapatite deposits at 12-14 days. NTPPPH activity increased steadily over 14 days. Transforming growth factor-beta and 1,25-dihydroxyvitamin D-3 increased PC-1 and NTPPPH in matrix vesicles (MVs), Because PC-1/NTPPPH was regulated in mineralizing MC3T3 cells, we stably transfected or infected cells with recombinant adenovirus, in order to express 2- to 6-fold more PC-1. PC-1/NTPPPH and PPi content increased severalfold in MVs derived from cells transfected with PC-1, Furthermore, MC3T3 cells transfected with PC-1 deposited similar to 80-90% less hydroxyapatite (by weight) than cells transfected with empty plasmid or enzymatically inactive PC-1, ATP-dependent Ca-45 precipitation by MVs from cells overexpressing active PC-1 was comparably diminished. Thus, regulation of PC-1 controls the PPi content and function of osteoblast-derived MVs and matrix hydroxyapatite deposition. PC-1 may provide a novel therapeutic target in certain disorders of bone mineralization,