PELP1 通过 ERK 通路促进人牙周膜干细胞成骨分化过程中 RUNX2 的表达。

PELP1 promotes the expression of RUNX2 via the ERK pathway during the osteogenic differentiation of human periodontal ligament stem cells.

机构信息

Department of Endodontics, Beijing Stomatological Hospital, School of Stomatology, Capital Medical University, Beijing, 10050, China.

Bejing Zebro Dental Clinic, Beijing, 100097, China.

出版信息

Arch Oral Biol. 2021 Apr;124:105078. doi: 10.1016/j.archoralbio.2021.105078. Epub 2021 Feb 11.

DOI:10.1016/j.archoralbio.2021.105078

PMID:33607589

Abstract

OBJECTIVE

The aim of this study was to determine the physiological function and mechanism of proline-, glutamic acid-, and leucine-rich protein 1 (PELP1) in the osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) at the molecular level in vitro.

DESIGN

During the osteogenic differentiation of hPDLSCs, the change of PELP1 and the osteogenic commitment markers runt-related transcription factor 2(RUNX2), alkaline phosphatase (ALP) and osteocalcin (OCN) were monitored by quantitative real-time PCR (qRT-PCR) and western blots. To elucidate how PELP1 regulates RUNX2, the expression of RUNX2, the phosphorylation of extracellular regulated protein kinases (ERK) and subcellular location of PELP1 were detected under conditions that PELP1 was either knockdown by specific siRNA or overexpressed. A pharmacological inhibitor of ERK, U0126 was used while PELP1 was overexpressed, and the expression of RUNX2 was monitored by qRT-PCR.

RESULTS

PELP1 was upregulated during the osteogenic differentiation of hPDLSCs. Knockdown of PELP1 suppressed the expression of RUNX2, whereas overexpression of PELP1 increased RUNX2 expression. Moreover, PELP1 knockdown resulted in reduced ERK phosphorylation and RUNX2 expression, and PELP1 overexpression induced RUNX2 expression was inhibited by U0126 in the hPDLSCs.

CONCLUSIONS

PELP1 regulates the expression of RUNX2 during the osteogenic differentiation of hPDLSCs and that the ERK pathway is involved in this process.

摘要

目的

本研究旨在从分子水平上探讨脯氨酸、谷氨酸和亮氨酸丰富蛋白 1（PELP1）在人牙周膜干细胞（hPDLSCs）成骨分化过程中的生理功能和作用机制。

设计

在 hPDLSCs 成骨分化过程中，通过实时定量 PCR（qRT-PCR）和 Western blot 监测 PELP1 与成骨定向标志物 runt 相关转录因子 2（RUNX2）、碱性磷酸酶（ALP）和骨钙素（OCN）的变化。为阐明 PELP1 如何调节 RUNX2，在 PELP1 被特异性 siRNA 敲低或过表达的情况下，检测 RUNX2 的表达、细胞外调节蛋白激酶（ERK）的磷酸化和 PELP1 的亚细胞定位。当 PELP1 过表达时，使用 ERK 的药理学抑制剂 U0126 来监测 RUNX2 的表达。

结果

在 hPDLSCs 的成骨分化过程中，PELP1 上调。PELP1 敲低抑制了 RUNX2 的表达，而过表达 PELP1 则增加了 RUNX2 的表达。此外，PELP1 敲低导致 ERK 磷酸化和 RUNX2 表达减少，而 U0126 抑制了 PELP1 过表达诱导的 hPDLSCs 中 RUNX2 的表达。