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PERK-eIF2α-ATF4 signaling contributes to osteogenic differentiation of periodontal ligament stem cells.
Journal of Molecular Histology ( IF 3.2 ) Pub Date : 2020-03-02 , DOI: 10.1007/s10735-020-09863-y Shuangyan Yang 1 , Lihua Hu 2 , Chunling Wang 1 , Fulan Wei 1
Journal of Molecular Histology ( IF 3.2 ) Pub Date : 2020-03-02 , DOI: 10.1007/s10735-020-09863-y Shuangyan Yang 1 , Lihua Hu 2 , Chunling Wang 1 , Fulan Wei 1
Affiliation
Protein kinase-like endoplasmic reticulum kinase (PERK) is a type I transmembrane protein located in the endoplasmic reticulum (ER). The PERK-eukaryotic initiation factor 2α (eIF2α)-activating transcription factor 4 (ATF4) pathway has been proved to be involved in osteoblast differentiation, but the involvement of the PERK-eIF2α-ATF4 signaling pathway in osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) has remained unclear. Therefore, the aim of this study was to explore the role of PERK in osteogenic differentiation of hPDLSCs and to assess whether PERK-eIF2α-ATF4 contributes to the process of osteogenic differentiation in hPDLSCs. In our study, we constructed PERK-overexpressed and PERK-silenced hPDLSCs by lentiviral transduction. Furthermore, lentivirus-transfected cells were induced to differentiate into osteoblast cells for different days. Alkaline phosphatase (ALP) activity and Alizarin Red staining were used to evaluate the mineralization capacity, and the expression levels of related genes-ATF4, ALP, bone sialoprotein, runt-related transcription factor 2 (Runx2), and osteocalcin were measured to evaluate the osteogenic differentiation of hPDLSCs. The results showed that over-expression of PERK greatly increased ALP activity and the expression levels of related osteogenic genes, which displayed the strongest osteogenesis capacity. However, suppression of PERK caused decreased ALP activity and the weakest osteogenesis capacity, and the levels of ATF4 and p-eIF2α in PERK-silenced hPDLSCs were also decreased. Our results indicated that the PERK gene plays an important role in the differentiation of hPDLSCs to osteoblast-like cells. The PERK-eIF2α-ATF4 signaling pathway contributes to osteoblast differentiation of hPDLSCs.
中文翻译:
PERK-eIF2α-ATF4信号传导有助于牙周膜干细胞的成骨分化。
蛋白激酶样内质网激酶(PERK)是位于内质网(ER)中的I型跨膜蛋白。已经证明PERK真核起始因子2α(eIF2α)激活转录因子4(ATF4)通路与成骨细胞分化有关,但PERK-eIF2α-ATF4信号通路与人牙周膜干细胞成骨分化有关。 (hPDLSC)仍不清楚。因此,本研究的目的是探讨PERK在hPDLSC的成骨分化中的作用,并评估PERK-eIF2α-ATF4是否有助于hPDLSC的成骨分化。在我们的研究中,我们通过慢病毒转导构建了PERK过表达和PERK沉默的hPDLSC。此外,慢病毒转染的细胞被诱导分化成成骨细胞不同天。用碱性磷酸酶(ALP)活性和茜素红染色评估矿化能力,并测量相关基因-ATF4,ALP,骨唾液蛋白,矮子相关转录因子2(Runx2)和骨钙蛋白的表达水平以评估矿化能力。 hPDLSC的成骨分化。结果表明,PERK的过表达大大提高了ALP活性和相关成骨基因的表达水平,显示出最强的成骨能力。然而,抑制PERK会导致ALP活性降低和最弱的成骨能力,并且PERK沉默的hPDLSCs中的ATF4和p-eIF2α的水平也会降低。我们的结果表明,PERK基因在hPDLSCs向成骨细胞样细胞的分化中起重要作用。PERK-eIF2α-ATF4信号通路有助于hPDLSC的成骨细胞分化。
更新日期:2020-03-02
中文翻译:
PERK-eIF2α-ATF4信号传导有助于牙周膜干细胞的成骨分化。
蛋白激酶样内质网激酶(PERK)是位于内质网(ER)中的I型跨膜蛋白。已经证明PERK真核起始因子2α(eIF2α)激活转录因子4(ATF4)通路与成骨细胞分化有关,但PERK-eIF2α-ATF4信号通路与人牙周膜干细胞成骨分化有关。 (hPDLSC)仍不清楚。因此,本研究的目的是探讨PERK在hPDLSC的成骨分化中的作用,并评估PERK-eIF2α-ATF4是否有助于hPDLSC的成骨分化。在我们的研究中,我们通过慢病毒转导构建了PERK过表达和PERK沉默的hPDLSC。此外,慢病毒转染的细胞被诱导分化成成骨细胞不同天。用碱性磷酸酶(ALP)活性和茜素红染色评估矿化能力,并测量相关基因-ATF4,ALP,骨唾液蛋白,矮子相关转录因子2(Runx2)和骨钙蛋白的表达水平以评估矿化能力。 hPDLSC的成骨分化。结果表明,PERK的过表达大大提高了ALP活性和相关成骨基因的表达水平,显示出最强的成骨能力。然而,抑制PERK会导致ALP活性降低和最弱的成骨能力,并且PERK沉默的hPDLSCs中的ATF4和p-eIF2α的水平也会降低。我们的结果表明,PERK基因在hPDLSCs向成骨细胞样细胞的分化中起重要作用。PERK-eIF2α-ATF4信号通路有助于hPDLSC的成骨细胞分化。