GINS2 affects activity/differentiation, apoptosis and proliferation of osteoblast and osteoclast in steroid-induced osteonecrosis of the femoral head by regulating P53/GADD45A signaling pathway

Lei Tian,Jian Wang,Wei Li,Xianquan Wang,Shui Sun

doi:10.1590/fst.09921

Abstract

To investigate the function and mechanism of GINS complex subunit 2 (GINS2) in osteoblast and osteoclast in steroid-induced osteonecrosis of the femoral head (SINFH). Clinical specimens were collected from 40 cases of SINFH patients and 40 cases of control patients. Then, SINFH mouse model was built. Immediately, osteoblasts and osteoclasts were induced and differented from the pre-osteoblast cell line MC3T3-E1 and the mononuclear macrophage cell line RAW264.7 of mice, respectively. The qRT-PCR assay was used for detecting the expressions of GINS2, p53 and growth arrest and DNA damage 45A (GADD45A). The cytoactive, proliferation and apoptosis of osteoblasts and osteoclasts of SINFH mice were measured by tartaric acid phosphatase staining, CCK-8 and Annexin V-FITC/PI dual staining, respectively. GINS2 was up-regulated while p53/GADD45A was down-regulated in femoral tissues in SINFH patients and mice. GINS2 was down-regulated while p53 and GADD45A were up-regulated in osteoblasts of SINFH mice, which presented opposite trends in osteoclasts formations. GINS2 decreased osteoblasts activities and increased osteoclasts activities, inhibited apoptosis and promoted proliferation in osteoblasts and osteoclasts via p53/GADD45A pathway. GINS2 affects activity/differentiation, apoptosis and proliferation of osteoblast and osteoclast in steroid-induced osteonecrosis of the femoral head by regulating p53/GADD45A signaling pathway.

Highlights

Osteonecrosis of the femoral head (ONFH) is a common refractory disease in the department of orthopedics leading to femoral head collapse and hip dyskinesia (Wang et al, 2019; Seixas et al, 2020)
3.1 The expressions of GINS complex subunit 2 (GINS2), p53 and growth arrest and DNA damage 45A (GADD45A) in clinical specimens quantitative reverse transcription-polymerase chain reaction (qRT-PCR) results showed that GINS2 was up-regulated while p53 and GADD45A were down-regulated in steroid-induced ONFH (SINFH) clinical specimens compared to the control tissues (P
3.2 The expressions of GINS2, p53 and GADD45A in femoral tissues of mice qRT-PCR results displayed that GINS2 was up-regulated while p53 and GADD45A were down-regulated in femoral tissues of SINFH model mice compared to the control group (P

Summary

Introduction

ONFH includes two types: non-traumatic ONFH and traumatic ONFH (Amanatullah et al, 2011; Mont et al, 2006). High incidence of non-traumatic ONFH is occurred in young and middle patients who aged 30-50 years, and the pathogeny of non-traumatic ONFH is multifactorial including hereditary susceptibility and risk factors for morbidity such as prolonged intensive use of hormones, alcohol, connective tissue disease, or hematologic illness (Amanatullah et al, 2011; Mont et al, 2006; Chen et al, 2019). Steroid-induced ONFH (SINFH), as an iatrogenic disease, the incidence of which is increasing with the years and usually develops at an early phase in the procedure of steroid administration (Zhang et al, 2013; Yu et al, 2019). Thereby, further exploring the pathogenesis of SINFH to look for new therapeutic targets of it is very urgent

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