Gene Expression and Protein Production of Leukemia Inhibitory Factor in Human Dental Follicle Cells
Gene Expression and Protein Production of Leukemia Inhibitory Factor in Human Dental Follicle Cells
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DOI：
10.5466/ijoms.11.156
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发表时间：
2013
期刊：
影响因子：
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通讯作者：
Y. Fujimoto;N. Ogura;H. Aonuma;Kosuke Takahashi;Ko Ito;T. Kondoh
中科院分区：
文献类型：
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作者：
Y. Fujimoto;N. Ogura;H. Aonuma;Kosuke Takahashi;Ko Ito;T. Kondoh
牙囊是一种外胚间充质组织，包围着发育中的牙胚，含有成骨/成牙骨质细胞谱系的干细胞/祖细胞。本研究的目的是检测白血病抑制因子(LIF)在人牙囊细胞(HDFC)中的基因表达和蛋白合成，并通过miRNA靶向在转录后水平对其进行调控。用实时荧光定量聚合酶链式反应检测骨髓间充质干细胞成骨诱导液(MSCOIM)培养0、1、2、4、7、11天HDFC中LIF基因的动态表达。MSCOIM培养中的表达呈时间依赖性下降。MSCOIM培养第2天LIF蛋白水平下降，第4天LIF蛋白水平升高。这些结果表明，LIF的表达在转录后水平上受到调控，并可能受到microRNA(MiRNA)的影响。为了证明特定的miRNAs在调节LIF产生中的直接作用，我们用四个预测靶向LIF的miRNAs的miRNA模拟物：hsa-miR-29b、hsa-miR-125a、hsa-miR-1995p和hsa-miR-199-3p转染HDFC。转染后，hsa-miR-125a、hsa-miR-199-5p和hsamiR-199-3p对LIF的产生无明显影响。这些结果表明，LIF的产生部分受miRNA的调节，维持HDFC的未分化状态，并影响向成骨/成牙骨质细胞系的分化。
The dental follicle is an ectomesenchymal tissue that surrounds developing tooth germs, which contains osteoblastic/cementoblastic lineage committed stem/progenitor cells. The purpose of this study is to examine the gene expression and the protein production of leukemia inhibitory factor(LIF)in human dental follicle cells(hDFC)and to regulate these at the post-transcriptional level by miRNA targeting. We examined the kinetic gene expression of LIF in hDFC cultured with mesenchymal stem cell osteogenic induction medium(MSCOIM)on days 0, 1, 2, 4, 7 and 11 using real-time PCR. Expression was decreased in MSCOIM culture in a time-dependent manner. LIF protein levels decreased until culture day 2, and then increased until day 4 in MSCOIM culture. These results suggest that LIF expression is regulated at the post-transcriptional level, and may be affected by microRNA(miRNA). To demonstrate the direct role of specific miRNAs in the modulation of LIF production, we transfected hDFC with miRNA mimics of four miRNAs predicted to target LIF; hsa-miR-29b, hsa-miR-125a, hsa-miR-1995p and hsa-miR-199-3p. After transfection, the production of LIF was significantly lower in the cells transfected with miRNA his-miR-29b, but hsa-miR-125a, hsa-miR-199-5p and hsamiR-199-3p showed no significant changes in LIF production. These results suggest that production of LIF is partially regulated by miRNA, maintaining the undifferentiated status of hDFC and influencing differentiation toward the osteoblastic/ cementoblastic lineage.