The Role of Integrin-α5 in the Proliferation and Odontogenic Differentiation of Human Dental Pulp Stem Cells
The Role of Integrin-α5 in the Proliferation and Odontogenic Differentiation of Human Dental Pulp Stem Cells
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DOI：
10.1016/j.joen.2013.08.011
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发表时间：
2014-02-01
影响因子：
4.2
通讯作者：
Wu, Buling
中科院分区：
文献类型：
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作者：
Cui, Li;Xu, Shuaimei;Wu, Buling
导语：已有报道称整合素- α 5 （ITGA5）活性与细胞增殖、分化、迁移和器官发育有关。然而，ITGA5在人牙髓干细胞（hDPSCs）生物学功能中的作用尚未被探索。本研究旨在探讨ITGA5在hdpsc增殖和成牙分化中的作用。方法：利用慢病毒介导的ITGA5短发夹RNA （shRNA）敲除hDPSCs中的ITGA5。采用3-（4,5-二甲基噻唑-2-基）-2,5-二苯基溴化四唑（MTT）测定和5-乙基-2'-脱氧尿苷标记法检测表达itga5特异性shRNA或阴性对照shRNA的慢病毒感染hDPSCs的增殖变化。将ITGA5敲除细胞和shMock细胞在矿化培养基中培养3周，用茜素红S染色检测细胞分化情况。采用实时聚合酶链反应和Western blot检测牙源性分化相关分子标志物的表达。结果：ITGA5基因的表达降低了hDPSCs的增殖能力。ITGA5 shRNA促进hdpsc的成牙分化，促进矿化结节的形成。上调多种牙本质生成标志物信使RNA的表达和牙本质唾液磷蛋白的表达。结论：这些发现提示ITGA5在维持hdpsc的增殖状态中起重要作用。抑制ITGA5信号通路可促进hdpsc向牙源性分化。
Introduction: It has been reported that integrin-alpha 5 (ITGA5) activity is related to cell proliferation, differentiation, migration, and organ development. However, the involvement of ITGA5 in the biological functions of human dental pulp stem cells (hDPSCs) has not been explored. The aim of this study was to investigate the role of ITGA5 in the proliferation and odontogenic differentiation of hDPSCs. Methods: We knocked down ITGA5 in hDPSCs using lentivirus-mediated ITGA5 short hairpin RNA (shRNA). Changes in the proliferation in hDPSCs infected with lentiviruses expressing ITGA5-specific shRNA or negative control shRNA were examined using the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and 5-ethynyl-2'-deoxyuridine labeling. Both ITGA5 knockdown cells and shMock cells were cultured in mineralization medium for 3 weeks, and the differentiation of cells was detected with alizarin red S staining. The expression of odontogenic differentiation-related molecular markers was assessed using real-time polymerase chain reaction and Western blot assays. Results: The knockdown of ITGA5 decreased the proliferation capacity of hDPSCs. ITGA5 shRNA promoted odontogenic differentiation of hDPSCs with the enhanced formation of mineralized nodules. It also up-regulated the messenger RNA expression of multiple markers of odontogenesis and the expression of dentin sialophosphoprotein protein. Conclusions: These findings suggest that ITGA5 plays an important role in maintaining hDPSCs in a proliferative state. The inhibition of ITGA5 signaling promotes the odontogenic differentiation of hDPSCs.