EGR1通过激活lnc-HCG11/Wnt/β-catenin通路促进牙周膜细胞的成骨分化

doi:10.12439/kqhm.1005-4979.2025.02.005

摘要/Abstract

摘要：

目的： 探讨早期生长应答因子1（early growth response gene 1，EGR1）是否通过调控长链非编码RNA人白细胞抗原复合物11（long non-coding RNA human leukocyte antigen complex group 11，lnc-HCG11）和无翅基因/β-连环蛋白（wingless/β-catenin，Wnt/β-catenin）通路影响人牙周膜干细胞（human periodontal ligament stem cells，hPDLSCs）的成骨分化。方法： 通过载体转染过表达和干扰EGR1和lnc-HCG11在hPDLSCs中的表达，通过β-catenin抑制剂IWR-1抑制该通路的活化水平，实时定量聚合酶链反应（real-time quantitative polymerase chain reaction，RT-qPCR）与蛋白质印迹法检测干预效果。对上述处理的hPDLSCs进行成骨诱导后，利用RT-qPCR检测成骨基因Runt相关转录因子2（runt-related transcription factor 2，RUNX2）、Ⅰ型胶原α1（collagen typeⅠα1，COL1A1）、骨桥蛋白（osteopontin，OPN）、骨钙素（osteocalcin，OCN）的表达水平，茜素红染色检测细胞的矿化水平。结果： 过表达EGR1促进了hPDLSCs中lnc-HCG11的表达，过表达lnc-HCG11促进hPDLSCs中RUNX2、COL1A1、OPN、OCN的表达水平及矿化水平，沉默lnc-HCG11抑制EGR1的促成骨分化作用，加入IWR-1后抑制了活性β-catenin的表达，同时削弱了lnc-HCG11对hPDLSCs的促成骨分化作用。结论： EGR1通过激活lnc-HCG11/Wnt/β-catenin通路促进hPDLSCs的成骨分化。

关键词: 牙周炎, 人牙周膜干细胞, 早期生长应答因子1, lncRNA HCG11, 成骨分化

Abstract:

Objective: To investigate the effect of early growth response gene 1 (EGR1) on osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) by regulating long non-coding RNA human leukocyte antigen complex group 11 (lnc-HCG11) and wingless/β-catenin (Wnt/β-catenin) pathway.Methods: The expression levels of EGR1 and lnc-HCG11 were changed in hPDLSCs through gene overexpression and interference assays, while the activation level of the Wnt/β-catenin pathway was altered by the β-catenin inhibitor IWR-1. Subsequently, real-time quantitative polymerase chain reaction (RT-qPCR) and Western blotting experiments were conducted to validate the intervention effect. After osteogenic induction, RT-qPCR was used to detect the expression levels of osteogenic genes runt-related transcription factor 2 (RUNX2), collagen type Ⅰα 1 (COL1A1), osteopontin (OPN), osteocalcin (OCN), and alizarin red staining was used to detect the mineralization level of cells.Results: The overexpression of EGR1 was observed to promote the expression of lnc-HCG11 in hPDLSCs, while the overexpression of lnc-HCG11 promoted the expression levels of RUNX2, COL1A1, OPN, OCN, and mineralization in hPDLSCs. The pro-osteogenic effect of EGR1 was inhibited by silencing lnc-HCG11, while treatment with IWR-1 suppressed the expression of active β-catenin and weakened the pro-osteogenic effect of lnc-HCG11 on hPDLSCs.Conclusion: EGR1 promotes osteogenic differentiation of hPDLSCs by activating the lnc-HCG11/Wnt/β-catenin pathway.

Key words: periodontitis, human periodontal ligament stem cells, early growth response gene 1, lncRNA HCG11, osteogenic differentiation

中图分类号:

R782

李成, 隆湘凤, 周琴, 徐方方, 黄学成, 毛加团. EGR1通过激活lnc-HCG11/Wnt/β-catenin通路促进牙周膜细胞的成骨分化[J]. 《口腔颌面外科杂志》, 2025, 35(2): 116-122.

LI Cheng, LONG Xiangfeng, ZHOU Qin, XU Fangfang, HUANG Xuecheng, MAO Jiatuan. EGR1 promotes osteogenic differentiation of periodontal ligament cells by activating the lnc-HCG11/Wnt/β-catenin pathway[J]. 《Journal of Oral and Maxillofacial Surgery》, 2025, 35(2): 116-122.

https://journal06.magtech.org.cn/Jweb_joms/CN/Y2025/V35/I2/116

图/表 4

图1 过表达EGR1促进了hPDLSCs中lnc-HCG11的表达 A. EGR1的mRNA相对表达水平；B. EGR1蛋白的表达条带；C. lnc-HCG11的mRNA表达水平。*表示P<0.05，与NC组比较。

Figure 1 Overexpression of EGR1 promoted the expression of lnc-HCG11 in hPDLSCs

图2 过表达lnc-HCG11促进hPDLSCs的成骨分化 A. 2组细胞lnc-HCG11的mRNA相对表达水平；B. 2组细胞中成骨相关基因RUNX2、COL1A1、OPN、OCN的mRNA相对表达水平；C、D. 2组细胞的矿化染色图及矿化水平统计结果。*表示P<0.05，与NC组比较。

Figure 2 Overexpression of lnc-HCG11 promoted osteogenic differentiation of hPDLSCs

图3 沉默lnc-HCG11抑制EGR1的促成骨分化作用 A. 各组细胞中EGR1的mRNA相对表达水平；B. 各组细胞中lnc-HCG11的mRNA相对表达水平；C. 各组细胞中RUNX2、COL1A1、OPN、OCN的mRNA相对表达水平；D、E. 各组细胞的矿化染色图及矿化水平统计结果。*表示P<0.05，与EGR1过表达组比较。

Figure 3 Silencing lnc-HCG11 inhibited the pro-osteogenic differentiation effect of EGR1

图4 抑制Wnt/β-catenin通路削弱lnc-HCG11的促成骨分化作用 A. 各组细胞活性β-catenin与总β-catenin的蛋白表达条带图；B. 各组细胞中RUNX2、COL1A1、OPN、OCN的mRNA相对表达水平；C、D. 各组细胞的矿化染色图及矿化水平统计结果。*表示P<0.05，与lnc-HCG11过表达组比较。

Figure 4 Inhibition of Wnt/β-catenin pathway weakened the pro-osteogenic differentiation effect of lnc-HCG11

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