Canfield A E, Doherty M J, Kelly V, Newman B, Farrington C, Grant M E, Boot-Handford R P
Wellcome Trust Centre for Cell-Matrix Research, School of Medicine, University of Manchester, Manchester, UK.
FEBS Lett. 2000 Dec 29;487(2):267-71. doi: 10.1016/s0014-5793(00)02363-2.
PCR-based subtractive hybridisation was used to identify genes up-regulated when pericytes undergo osteogenic differentiation and deposit a calcified matrix. cDNA pools were generated from confluent pericytes and from pericyte cultures containing calcified nodules. A pericyte cDNA library was screened with the product of the subtraction procedure (calcified minus confluent cDNA) and the majority of the positive clones were identified as matrix Gla protein (MGP). Northern analysis and immunohistochemistry demonstrated that MGP was only expressed by pericytes in calcified nodules. Antibodies to MGP inhibited the deposition of a calcified matrix by pericytes, suggesting that MGP regulates both cell differentiation and calcification.
基于聚合酶链反应（PCR）的消减杂交技术被用于鉴定在周细胞发生成骨分化并沉积钙化基质时上调的基因。从汇合的周细胞以及含有钙化结节的周细胞培养物中生成互补DNA（cDNA）文库。用消减程序的产物（钙化的减去汇合的cDNA）筛选周细胞cDNA文库，大多数阳性克隆被鉴定为基质γ-羧基谷氨酸蛋白（MGP）。Northern印迹分析和免疫组织化学表明，MGP仅在钙化结节中的周细胞中表达。抗MGP抗体抑制了周细胞对钙化基质的沉积，这表明MGP调节细胞分化和钙化。
