Department of Maxillofacial Tissue Regeneration, School of Dentistry, Kyung Hee University, Seoul, South Korea.
J Endod. 2012 Jun;38(6):769-73. doi: 10.1016/j.joen.2012.03.014. Epub 2012 Apr 21.
The lysyl oxidase (LOX) family is an emerging family of amine oxidases responsible for the formation of collagen fibrils in the extracellular matrix. To date, 5 LOX family genes have been identified in humans, encoding LOX and LOX-like proteins (LOXL, LOXL2, LOXL3, and LOXL4). The goal of this study was to evaluate the expression and function of the LOX family genes in odontoblastic differentiation of human dental pulp (HDP) cells.
Expression of the LOX family genes was assessed by reverse transcriptase polymerase chain reaction analysis, and the amine oxidase activity of HDP cells was evaluated by peroxidase-coupled fluorometric assays. Mineral nodule formation and expression of odontoblastic marker genes were assessed in the presence and absence of specific small interfering RNAs (siRNAs) of the LOX family genes.
Among the LOX family genes, only LOX and LOXL showed prominent expression during odontoblastic differentiation of HDP cells. Suppression of LOX and LOXL expression by siRNA-induced interference substantially decreased the amine oxidase activity of the differentiating HDP cells. Furthermore, interference of LOX and LOXL expression inhibited mineral nodule formation and expression of odontoblastic marker genes during odontoblastic differentiation of HDP cells.
These findings show for the first time that the LOX- and LOXL-mediated organization of collagen fibrils in extracellular matrices of HDP cells might be an important regulator for odontoblastic differentiation of HDP cells.
赖氨酰氧化酶(LOX)家族是一个新兴的胺氧化酶家族,负责细胞外基质中胶原纤维的形成。迄今为止,人类已经鉴定出 5 种 LOX 家族基因,分别编码 LOX 和 LOX 样蛋白(LOXL、LOXL2、LOXL3 和 LOXL4)。本研究的目的是评估 LOX 家族基因在人牙髓(HDP)细胞成牙本质分化中的表达和功能。
通过逆转录聚合酶链反应分析评估 LOX 家族基因的表达,并通过过氧化物酶偶联荧光测定评估 HDP 细胞的胺氧化酶活性。在存在和不存在 LOX 家族基因的特异性小干扰 RNA(siRNA)的情况下,评估矿化结节的形成和牙本质标志物基因的表达。
在 LOX 家族基因中,只有 LOX 和 LOXL 在 HDP 细胞的成牙本质分化过程中表现出明显的表达。siRNA 诱导干扰 LOX 和 LOXL 的表达显著降低了分化中的 HDP 细胞的胺氧化酶活性。此外,干扰 LOX 和 LOXL 的表达抑制了矿化结节的形成和牙本质标志物基因在 HDP 细胞成牙本质分化过程中的表达。
这些发现首次表明,LOX 和 LOXL 介导的 HDP 细胞细胞外基质中胶原纤维的排列可能是 HDP 细胞成牙本质分化的重要调节因子。