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Effects of GPNMB on proliferation and odontoblastic differentiation of human dental pulp cells

机译:GPNMB对人牙髓细胞增殖和成牙本质细胞分化的影响

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摘要

Glycoprotein (transmembrane) nonmetastatic melanoma protein b (GPNMB) plays crucial roles in odontogenesis. However, the role of GPNMB in human dental pulp cells (hDPCs) is still unclear. Therefore, in this study, we investigated the expression and function of the GPNMB in odontoblastic differentiation of hDPCs. Cells were cultured in odontoblast differentiation-inducing medium; the expression of the GPNMB was assessed by reverse transcriptase polymerase chain reaction and Western blot analysis. We performed gene knockdown of GPNMB in hDPCs using lentivirus-mediated small interfering RNA (siRNA)-GPNMB. The proliferation of cells was measured by the MTT assay, and the differentiation of cells was detected with alkaline phosphatase (ALP) activity assay, qRT-PCR and Western blot were used to determine the expression levels of dentin sialophosphoprotein (DSPP) and dentin matrix protein-1 (DMP-1). The expression level of GPNMB was significantly increased during odontoblastic differentiation of hDPCs. Suppression of GPNMB expression by siRNA-GPNMB obviously promoted the proliferation of hDPCs. Furthermore, siRNA-GPNMB significantly inhibited the activity of ALP and expression levels of DSPP and DMP-1 during odontoblastic differentiation of hDPCs. Our results show that GPNMB plays an important role in regulating the expression of key pluripotency genes in hDPCs and modifying odontogenic differentiation.
机译:糖蛋白(跨膜)非转移性黑色素瘤蛋白b(GPNMB)在牙生成中起关键作用。但是,GPNMB在人类牙髓细胞(hDPC)中的作用仍不清楚。因此,在这项研究中,我们调查了GPNMB在hDPC的牙本质成骨分化中的表达和功能。细胞在成牙本质细胞分化诱导培养基中培养。通过逆转录酶聚合酶链反应和蛋白质印迹分析评估GPNMB的表达。我们使用慢病毒介导的小干扰RNA(siRNA)-GPNMB在hDPC中进行了GPNMB的基因敲低。 MTT法检测细胞的增殖,碱性磷酸酶(ALP)活性法检测细胞的分化,qRT-PCR和Western blot检测牙本质唾液磷蛋白(DSPP)和牙本质基质蛋白的表达水平。 -1(DMP-1)。在hDPCs的成牙本质细胞分化过程中,GPNMB的表达水平显着增加。 siRNA-GPNMB抑制GPNMB表达明显促进了hDPCs的增殖。此外,siRNA-GPNMB在hDPC的成牙细胞分化过程中显着抑制了ALP的活性以及DSPP和DMP-1的表达水平。我们的结果表明,GPNMB在调节hDPC中关键多能性基因的表达和修饰牙源性分化中起着重要作用。

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