Understanding the functional role of genistein in the bone differentiation in mouse osteoblastic cell line MC3T3-E1 by RNA-seq analysis

Abstract

Genistein, a phyto-estrogen, can potentially replace endogenous estrogens in postmenopausal women, but the underlying molecular mechanisms remain incompletely understood. To obtain insight into the effect of genistein on bone differentiation, RNA sequencing (RNA-seq) analysis was used to detect differentially expressed genes (DEGs) in genistein-treated vs. untreated MC3T3-E1 mouse osteoblastic cells. Osteoblastic cell differentiation was monitored by measuring osteoblast differentiation factors (ALP production, bone mineralization, and expression of osteoblast differentiation markers). From RNA-seq analysis, a total of 132 DEGs (including 52 up-regulated and 80 down-regulated genes) were identified in genistein-treated cells (FDR q-value < 0.05 and fold change > 1.5). KEGG pathway and Gene Ontology (GO) enrichment analyses were performed to estimate the biological functions of DEGs and demonstrated that these DEGs were highly enriched in functions related to chemotactic cytokines. The functional relevance of DEGs to genistein-induced osteoblastic cell differentiation was further evaluated by siRNA-mediated knockdown in MC3T3-E1 cells. These siRNA knockdown experiments (of the DEGs validated by real-time qPCR) demonstrated that two up-regulated genes (Ereg and Efcab2) enhance osteoblastic cell differentiation, while three down-regulated genes (Hrc, Gli, and Ifitm5) suppress the differentiation. These results imply their major functional roles in bone differentiation regulated by genistein.

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“…55 In addition, genistein and daidzein also induce osteoblastic differentiation and angiogenesis on bone formation. [56][57][58] In the present study, Jix inhibited Pg infection and decreased alveolar bone resorption. Osteoblasts appear on tension side to promote new bone formation.…”

Flavonoids: Classification, Function, and Molecular Mechanisms Involved in Bone Remodelling

“…In MC3T3 pre-osteoblastic cells, treatment with genistein altered the expression levels of genes associated with cell proliferation, cell migration, cell differentiation, and inflammatory responses. Successive knockdown analyses showed that two upregulated genes (Ereg and Efcab2) and three downregulated genes (Hrc, Gli1, and Iftm5) play crucial roles in the differentiation of osteoblasts via increasing the expressions of osteoblast-associated markers such as RUNX2, ALP and BMP-2 ( 146 ). Administration of genistein to human bone marrow stromal cells suppressed its differentiation into adipocytes by inhibiting the mRNA levels of PPARg and CCAAT/enhancer binding proteins (C/EBPs), while it enhanced osteoblastogenesis, thus preventing bone loss associated with excessive adipogenesis ( 147 ).…”

Emerging Natural-Product-Based Treatments for the Management of Osteoarthritis

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