Lymphocyte enhancer-binding factor 1: an essential factor in odontoblastic differentiation of dental pulp cells enzymatically isolated from rat incisors
Lymphocyte enhancer-binding factor 1: an essential factor in odontoblastic differentiation of dental pulp cells enzymatically isolated from rat incisors
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DOI：
10.1007/s00774-010-0185-0
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发表时间：
2010-04
影响因子：
3.3
通讯作者：
S. Yokose;T. Naka
中科院分区：
文献类型：
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作者：
S. Yokose;T. Naka
淋巴细胞增强结合因子1 （Lef1）是一种hmg结构域蛋白，被认为在牙齿发育过程中诱导组织相互作用中起重要作用。在小鼠中，lef1基因敲除会导致牙齿发育停滞在萌芽阶段。由于该基因在牙乳头外充质细胞分化过程中参与调控大量多样的肽生长因子，因此lef1似乎是成牙细胞分化的关键因子。然而，lef1与成牙细胞分化的关系尚不清楚。为了分析lef1在调节成牙细胞分化中的生物学作用，我们在培养的牙髓细胞中短暂过表达或抑制lef1。与对照细胞相比，过表达lef1的细胞表达了更高水平的牙本质唾液蛋白（DSPP）、骨钙素和碱性磷酸酶（ALP） mRNA，形成了更多的矿化结节。然而，过表达lef1不影响msx -1的表达或细胞增殖。为了进一步研究lef1在牙髓细胞中的作用，我们使用短干扰RNA （siRNA）敲低了lef1在牙髓细胞中的表达。siRNA对lef1的瞬时表达显著降低了lef1mrna水平，与对照细胞相比，lef1抑制的细胞表达的dspp、骨钙素和alpmrna水平较低。此外，siRNA抗stlef1可抑制矿化结核的形成；然而，针对lef1的siRNA既不能抑制msx -1的表达，也不能抑制细胞增殖。这些结果概述了Lef1通过调节牙髓细胞中dspandosteocalcinmrna的表达来加速成牙细胞分化的作用，证实了Lef1是成牙细胞分化的关键因子。
Lymphocyte enhancer-binding factor 1 (Lef1), a HMG-domain protein, is thought to play important roles in inductive tissue interaction during tooth development.Lef1knockdown in mice causes arrest at the bud stage in tooth development. As this gene participates in the regulation of a large and diverse set of peptide growth factors in ectomesenchymal cell differentiation of dental papilla,Lef1appears to be a key factor in odontoblast differentiation. However, the relationship betweenLef1and odontoblast differentiation is still unclear. To analyze the biological roles ofLef1in regulating odontoblast differentiation, we transiently overexpressed or suppressedLef1in cultured dental pulp cells.Lef1-overexpressing cells expressed higher levels ofdentin sialoprotein(DSPP),osteocalcinandalkaline phosphatase(ALP) mRNA and formed larger numbers of mineralized nodules compared to control cells. However,Msx-1expression or cell proliferation was unaffected by overexpression ofLef1. To further examine the role ofLef1in dental pulp cells, we knocked downLef1expression in dental pulp cells using short interfering RNA (siRNA). Transient expression of siRNA againstLef1markedly reducedLef1mRNA levels, andLef1-suppressed cells expressed lower levels ofDSPP,osteocalcinandALPmRNA compared to control cells. Furthermore, the formation of mineralized nodules was inhibited by siRNA againstLef1; however, neitherMsx-1expression or cell proliferation was inhibited by siRNA againstLef1. These results outline the role ofLef1in accelerating odontoblast differentiation by regulatingDSPandosteocalcinmRNA expression in dental pulp cells, confirming that Lef1 is a key factor for odontoblast differentiation.