McLachlan Julia L, Smith Anthony J, Bujalska Iwona J, Cooper Paul R
Oral Biology, School of Dentistry, University of Birmingham, Birmingham, B4 6NN, UK.
Biochim Biophys Acta. 2005 Sep 25;1741(3):271-81. doi: 10.1016/j.bbadis.2005.03.007. Epub 2005 Apr 8.
High-throughput characterisation of the molecular response of pulpal tissue under carious lesions may contribute to improved future diagnosis and treatment. To identify genes associated with this process, oligonucleotide microarrays containing approximately 15,000 human sequences were screened using pooled total RNA isolated from pulpal tissue from both healthy and carious teeth. Data analysis identified 445 genes with 2-fold or greater difference in expression level, with 85 more abundant in health and 360 more abundant in disease. Subsequent gene ontological grouping identified a variety of processes and functions potentially activated or down-modulated during caries. Validation of microarray results was obtained by a combination of real-time and semi-quantitative PCR for selected genes, confirming down-regulation of Dentin Matrix Protein-1 (DMP-1), SLIT 2, Period-2 (PER 2), Period-3 (PER 3), osteoadherin, Glypican-3, Midkine, activin receptor interacting protein-1 (AIP 1), osteoadherin and growth hormone receptor (GHR), and up-regulation of Adrenomedullin (ADM), Interleukin-11 (IL-11), Bone sialoprotein (BSP), matrix Gla protein (MGP), endothelial cell growth factor-1 (ECGF 1), inhibin beta A and orosomucoid-1 (ORM 1), in diseased pulp. Real-time PCR analyses of ADM and DMP-1 in a panel of healthy and carious pulpal tissue and also in immune system cells highlighted the heterogeneity of caries and indicated increased expression of ADM in neutrophils activated by bacterial products. In contrast, DMP-1 was predominantly expressed by cells native to healthy pulpal tissue. This study has greatly extended our molecular knowledge of dental tissue disease and identified involvement of genes previously unassociated with this process.
高通量表征龋损下牙髓组织的分子反应可能有助于改善未来的诊断和治疗。为了鉴定与该过程相关的基因，使用从健康和龋损牙齿的牙髓组织中分离的混合总RNA，对包含约15,000个人类序列的寡核苷酸微阵列进行筛选。数据分析确定了445个基因，其表达水平差异为2倍或更大，其中85个在健康状态下表达更丰富，360个在疾病状态下表达更丰富。随后的基因本体分组确定了龋病过程中可能被激活或下调的各种过程和功能。通过对选定基因进行实时和半定量PCR相结合的方法，对微阵列结果进行了验证，证实了牙本质基质蛋白-1（DMP-1）、SLIT 2、周期蛋白-2（PER 2）、周期蛋白-3（PER 3）、骨黏附素、磷脂酰肌醇蛋白聚糖-3、中期因子、激活素受体相互作用蛋白-1（AIP 1）、骨黏附素和生长激素受体（GHR）在患病牙髓中下调，而肾上腺髓质素（ADM）、白细胞介素-11（IL-11）、骨唾液蛋白（BSP）、基质Gla蛋白（MGP）、内皮细胞生长因子-1（ECGF 1）、抑制素βA和类orosomucoid-1（ORM 1）上调。对一组健康和龋损牙髓组织以及免疫系统细胞中的ADM和DMP-1进行实时PCR分析，突出了龋病的异质性，并表明细菌产物激活的中性粒细胞中ADM表达增加。相比之下，DMP-1主要由健康牙髓组织中的固有细胞表达。这项研究极大地扩展了我们对牙组织疾病的分子认识，并确定了以前与该过程无关的基因的参与情况。
