State Key Laboratory of Oral Diseases, National Clinical Research Center for Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, China.
Stomatology Hospital, School of Stomatology, Zhejiang University School of Medicine, Zhejiang Provincial Clinical Research Center for Oral Diseases, Key Laboratory of Oral Biomedical Research of Zhejiang Province, Cancer Center of Zhejiang University, Hangzhou, China.
Cell Prolif. 2024 Feb;57(2):e13546. doi: 10.1111/cpr.13546. Epub 2023 Sep 20.
Scaffold protein AF4/FMR2 family member 4 (AFF4) has been found to play a role in osteogenic commitment of stem cells. However, function of AFF4 in human periodontal ligament stem cells (hPDLSCs) has not been studied yet. This present study aims to investigate the biological effect of AFF4 on osteogenic differentiation of hPDLSCs and potential mechanistic pathway. First, AFF4 expression profile was evaluated in conditions of periodontitis and osteogenic differentiation of hPDLSCs by immunohistochemical staining, western blot and qRT-PCR. Next, si-RNA mediated knockdown and lentiviral transduction mediated overexpression of AFF4 were adopted to explore impact of AFF4 on osteogenic capacity of hPDLSCs. Then, possible mechanistic pathway was identified. At last, pharmacological agonist of autophagy, rapamycin, was utilized to affirm the role of autophagy in AFF4-regulated osteogenesis of hPDLSCs. First, AFF4 expressions were significantly lower in inflamed periodontal tissues and lipopolysaccharides-treated hPDLSCs than controls, and were up-regulated during osteogenic differentiation of hPDLSCs. Next, osteogenic potential of hPDLSCs was impaired by AFF4 knockdown and potentiated by AFF4 overexpression. Moreover, AFF4 was found to positively regulate autophagic activity in hPDLSCs. At last, rapamycin treatment was shown to be able to partly restore AFF4 knockdown-suppressed osteogenic differentiation. Our study demonstrates that AFF4 regulates osteogenic potential of hPDLSCs via targeting autophagic activity. The involvement of AFF4 in periodontal homeostasis was identified for the first time.
支架蛋白 AF4/FMR2 家族成员 4（AFF4）已被发现在干细胞成骨分化中发挥作用。然而，AFF4 在人牙周膜干细胞（hPDLSCs）中的功能尚未研究。本研究旨在探讨 AFF4 对 hPDLSCs 成骨分化的生物学影响及其潜在的机制途径。首先，通过免疫组织化学染色、western blot 和 qRT-PCR 评估了 AFF4 在牙周炎和 hPDLSCs 成骨分化条件下的表达谱。接下来，采用 si-RNA 介导的敲低和慢病毒转导介导的 AFF4 过表达来探讨 AFF4 对 hPDLSCs 成骨能力的影响。然后，确定了可能的机制途径。最后，利用自噬的药理学激动剂雷帕霉素来证实自噬在 AFF4 调节 hPDLSCs 成骨中的作用。首先，与对照相比，在炎症性牙周组织和脂多糖处理的 hPDLSCs 中 AFF4 的表达明显降低，并且在 hPDLSCs 的成骨分化过程中上调。接下来，AFF4 敲低会损害 hPDLSCs 的成骨潜力，而过表达 AFF4 则增强其成骨潜力。此外，发现 AFF4 可正向调节 hPDLSCs 中的自噬活性。最后，雷帕霉素处理显示能够部分恢复 AFF4 敲低抑制的成骨分化。我们的研究表明，AFF4 通过靶向自噬活性来调节 hPDLSCs 的成骨潜力。首次确定了 AFF4 参与牙周稳态的作用。
