Oral Health Prev Dent. 2022 Apr 27;20:173-184. doi: 10.3290/j.ohpd.b2960495.
Recently, increasing attention has been paid to the function of long non-coding RNAs (lncRNAs) in osteogenic differentiation (OD) of dental pulp stem cells (DPSCs). LINC01133 was reported to have a close relationship with tumorigenesis for multiple cancers, but no study has yet explored the role of LINC01133 in modulating OD of DPSCs.
Alizarin red S (ARS) staining and alkaline phosphatase (ALP) staining were perfomed to assess the OD potential of DPSCs. Osteogenic markers including runt-related transcription factor 2 (RUNX2), osterix (OSX) and ALP expression levels in DPSCs were monitored by qRT-PCR and Western blot before and after cell transfection. Luciferase reporter gene assay detected the relationship between LINC01133 and miR-199b-5p.
The expression of LINC01133 was low, while miR-199b-5p was increasingly expressed during OD of DPSCs. Overexpression of LINC01133 in DPSCs resulted in decreased expression of RUNX2, OSX, ALP, DSPP and DMP1, whose expression was reversed in DPSCs after transfections of miR-199b-5p overexpression. Co-transfection of pcDNA3.1-LINC01133 and miR-199b-5p mimic led to elevated expression of RUNX2, OSX, ALP, DSPP and DMP1 compared with pcDNA3.1-LINC01133 transfection alone. LINC01133 served as a sponge of miR-199b-5p. AKT3 was verified as a downstream effector of miR-199b-5p in DPSCs.
LINC01133 inhibits the OD of DPSCs by upregulating AKT3 via sponging miR-199b-5p, which may act as a potential diagnostic biomarker for dentin regeneration in the dental pulp.
最近，人们越来越关注长链非编码 RNA（lncRNA）在牙髓干细胞（DPSC）成骨分化（OD）中的功能。LINC01133 被报道与多种癌症的发生密切相关，但尚无研究探讨 LINC01133 调节 DPSCs OD 的作用。
通过茜素红 S（ARS）染色和碱性磷酸酶（ALP）染色评估 DPSCs 的 OD 潜能。在细胞转染前后，通过 qRT-PCR 和 Western blot 监测 DPSCs 中成骨标志物 runt 相关转录因子 2（RUNX2）、osterix（OSX）和 ALP 的表达水平。荧光素酶报告基因检测 LINC01133 与 miR-199b-5p 的关系。
DPSC 成骨过程中 LINC01133 表达下调，miR-199b-5p 表达上调。在 DPSCs 中转染 LINC01133 过表达后，RUNX2、OSX、ALP、DSPP 和 DMP1 的表达降低，而过表达 miR-199b-5p 后则逆转了这一现象。与单独转染 pcDNA3.1-LINC01133 相比，共转染 pcDNA3.1-LINC01133 和 miR-199b-5p 模拟物可导致 RUNX2、OSX、ALP、DSPP 和 DMP1 的表达升高。LINC01133 是 miR-199b-5p 的海绵体。在 DPSCs 中，AKT3 被验证为 miR-199b-5p 的下游效应物。
LINC01133 通过海绵吸附 miR-199b-5p 上调 AKT3 抑制 DPSCs 的 OD，这可能成为牙髓再生的潜在诊断生物标志物。
