Varallo Vincenzo M, Gan Bing Siang, Seney Shannon, Ross Douglas C, Roth James H, Richards Robert S, McFarlane Robert M, Alman Benjamin, Howard Jeffrey C
Cell and Molecular Biology Laboratory, Hand & Upper Limb Centre, University of Western Ontario, London, Canada.
Oncogene. 2003 Jun 12;22(24):3680-4. doi: 10.1038/sj.onc.1206415.
Dupuytren's disease (DD) is a superficial fibromatosis of the hand. Although the molecular mechanisms responsible for this disease are unknown, recent studies suggest that beta-catenin may be a key factor involved in fibromatosis. In this study, we analysed the in vivo and in vitro expression levels of beta-catenin in DD, using surgical specimens and primary cell lines. Although no somatic mutations (exon 3) of beta-catenin were detected, Western blot analysis revealed high levels of beta-catenin in diseased palmar fascia, and low to undetectable levels of beta-catenin in patient-matched normal palmar fascia. Immunohistochemistry analysis showed high levels of beta-catenin expression within the disease fascia, as well as cytoplasmic and nuclear accumulations of the protein. Immunoprecipitation of beta-catenin from seven patient lesions showed the protein to be tyrosine phosphorylated. Lastly, Western analysis of three patient-matched (disease and normal fascia) primary cell cultures showed significantly elevated levels of beta-catenin in disease cells cultured in three-dimensional collagen lattices. This is the first extensive in vivo and in vitro characterization of beta-catenin in DD, and the first to suggest that the extracellular matrix may play an important role in modulating beta-catenin stability in DD.
掌腱膜挛缩症(DD)是一种手部的浅表纤维瘤病。尽管导致这种疾病的分子机制尚不清楚,但最近的研究表明,β-连环蛋白可能是参与纤维瘤病的关键因素。在本研究中,我们使用手术标本和原代细胞系分析了DD中β-连环蛋白的体内和体外表达水平。尽管未检测到β-连环蛋白的体细胞突变(外显子3),但蛋白质印迹分析显示,患病掌腱膜中β-连环蛋白水平较高,而与患者匹配的正常掌腱膜中β-连环蛋白水平低至无法检测到。免疫组织化学分析显示,疾病筋膜内β-连环蛋白表达水平较高,且该蛋白在细胞质和细胞核中积聚。对7例患者病变组织进行β-连环蛋白免疫沉淀显示该蛋白发生酪氨酸磷酸化。最后,对3例患者匹配的(疾病和正常筋膜)原代细胞培养物进行蛋白质印迹分析显示,在三维胶原晶格中培养的疾病细胞中β-连环蛋白水平显著升高。这是首次对DD中β-连环蛋白进行广泛的体内和体外特性分析,也是首次表明细胞外基质可能在调节DD中β-连环蛋白稳定性方面发挥重要作用。