Evaluation of the stability of reference genes in bone mesenchymal stem cells from patients with avascular necrosis of the femoral head

Wang, X N; Yang, Qiwei; Du, Zhenwu; Yu, Ting; Qin, Yunxia; Song, Yang; Xu, Mian; Wang, J C

doi:10.4238/gmr.15027926

Cited by 3 publications

(1 citation statement)

References 23 publications

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“…In 2008, TBP was found to be the most stable gene in the cartilage tissues of osteoarthritis patients [28]. Another researcher found PPIA to be the stable RG in the bone MSCs of patients with avascular necrosis of the femoral head [29]. Watanabe et al, recommended HPRT1, GAPDH, and YWHAZ RGs for gene analysis in the synovium of OA patients [30].…”

Section: Discussionmentioning

confidence: 99%

Insensitive Effects of Inflammatory Cytokines on the Reference Genes of Synovial Fluid Resident-Mesenchymal Stem Cells Derived from Rheumatoid Arthritis Patients

Bok,

Kim,

Thakur

et al. 2023

IJMS

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Mesenchymal stem cells derived from rheumatoid arthritis patients (RA-MSCs) provide an understanding of a variety of cellular and immunological responses within the inflammatory milieu. Sustained exposure of MSCs to inflammatory cytokines is likely to exert an influence on genetic variations, including reference genes (RGs). The sensitive effect of cytokines on the reference genes of RA-SF-MSCs may be a variation factor affecting patient-derived MSCs as well as the accuracy and reliability of data. Here, we comparatively evaluated the stability levels of nine RG candidates, namely GAPDH, ACTB, B2M, EEF1A1, TBP, RPLP0, PPIA, YWHAZ, and HPRT1, to find the most stable ones. Alteration of the RG expression was evaluated in MSCs derived from the SF of healthy donors (H-SF-MSCs) and in RA-SF-MSCs using the geNorm and NormFinder software programs. The results showed that TBP, PPIA, and YWHAZ were the most stable RGs for the normalization of H-SF-MSCs and RA-SF-MSCs using RT-qPCR, whereas ACTB, the most commonly used RG, was less stable and performed poorly. Additionally, the sensitivity of RG expression upon exposure to proinflammatory cytokines (TNF-α and IL-1β) was evaluated. RG stability was sensitive in the H-SF-MSCs exposed to TNF-α and IL-1β but insensitive in the RA-SF-MSCs. Furthermore, the normalization of IDO expression using ACTB falsely diminished the magnitude of biological significance, which was further confirmed with a functional analysis and an IDO activity assay. In conclusion, the results suggest that TBP, PPIA, and YWHAZ can be used in SF-MSCs, regardless of their exposure to inflammatory cytokines.

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Section: Discussionmentioning

confidence: 99%

Insensitive Effects of Inflammatory Cytokines on the Reference Genes of Synovial Fluid Resident-Mesenchymal Stem Cells Derived from Rheumatoid Arthritis Patients

Bok,

Kim,

Thakur

et al. 2023

IJMS

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The optimal compound reference genes for qRT-PCR analysis in the developing rat long bones under physiological conditions and prenatal dexamethasone exposure model

Han

Chen

et al. 2020

Reproductive Toxicology

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Differentially Expressed Genes Reveal the Biomarkers and Molecular Mechanism of Osteonecrosis

Zhang

Shi

2022

Journal of Healthcare Engineering

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Osteonecrosis is one of the most refractory orthopedic diseases, which seriously threatens the health of old patients. High-throughput sequencing (HTS) and microarray analysis have confirmed as an effective way for investigating the pathological mechanism of disease. In this study, GSE7716, GSE74089, and GSE123568 were obtained from Gene Expression Omnibus (GEO) database and used to identify differentially expressed genes (DEGs) by R language. Subsequently, the DEGs were analyzed with Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment. Moreover, the protein-protein interaction (PPI) network of DEGs was analyzed by STRING database and Cytoscape. The results showed that 318 downregulated genes and 58 upregulated genes were observed in GSE7116; 690 downregulated genes and 1148 upregulated genes were screened from 34183 genes in GSE74089. The DEGs involved in progression of osteonecrosis involved inflammation, immunological rejection, and bacterial infection-related pathways. The GO enrichment showed that osteonecrosis was related with extracellular matrix, external encapsulating structure organization, skeletal system development, immune response activity, cell apoptosis, mononuclear cell differentiation, and serine/threonine kinase activity. Moreover, PPI network showed that the progression of osteonecrosis of the femoral head was related with CCND1, CDH1, ESR1, SPP1, LOX, JUN, ITGA, ABL1, and VEGF, and osteonecrosis of the jaw is related with ACTB, CXCR4, PTPRC, IL1B, CXCL8, TNF, JUN, PTGS2, FOS, and RHOA. In conclusion, this study identified the hub factors and pathways which might play important roles in progression of osteonecrosis and could be used as potential biomarkers for diagnosis and treatment of osteonecrosis.

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Evaluation of the stability of reference genes in bone mesenchymal stem cells from patients with avascular necrosis of the femoral head

Cited by 3 publications

References 23 publications

Insensitive Effects of Inflammatory Cytokines on the Reference Genes of Synovial Fluid Resident-Mesenchymal Stem Cells Derived from Rheumatoid Arthritis Patients

Insensitive Effects of Inflammatory Cytokines on the Reference Genes of Synovial Fluid Resident-Mesenchymal Stem Cells Derived from Rheumatoid Arthritis Patients

The optimal compound reference genes for qRT-PCR analysis in the developing rat long bones under physiological conditions and prenatal dexamethasone exposure model

Differentially Expressed Genes Reveal the Biomarkers and Molecular Mechanism of Osteonecrosis

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