Let-7b downgrades CCND1 to repress osteogenic proliferation and differentiation of MC3T3-E1 cells: An implication in osteoporosis.
Abstract
The aim of this study was to reveal the effect of let-7b on osteoporosis (OP). Synthetic let-7b mimics or inhibitors were transfected into MC3T3-E1 cells. The expression of let-7b in MC3T3-E1 and its effect on cell viability, apoptosis, and the apoptosis-related proteins (Bcl-2, Bax, and cleaved caspase-9) were tested by CCK-8 assay, flow cytometry and Western blot, severally. The osteogenic differentiation markers (Runx2 and Osterix) and Wnt/β-catenin pathway related markers (β-catenin and C-myc) were detected by qRT-PCR and Western blot. The relationships between let-7b and cyclin D1 (CCND1) were confirmed by luciferase reporter assay. The differentiation and mineralization of MC3T3-E1 cells were analyzed by alkaline phosphatase (ALP) activity assay and alizarin red staining. The outcomes indicated that overexpression/ablation of let-7b repressed/facilitated MC3T3-E1 cell viability and accelerated/suppressed MC3T3-E1 cell apoptosis. Besides, a remarkable decrease/augment of Bcl-2 protein expression and the distinct fortify/reduction of Bax and cleaved caspase-9 expression levels were observed in let-7b mimics/inhibitors group in MC3T3-E1 cells. Moreover, we discovered that let-7b overexpression/ablation retrained/facilitated ...Continue Reading
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Apoptosis
Apoptosis is a specific process that leads to programmed cell death through the activation of an evolutionary conserved intracellular pathway leading to pathognomic cellular changes distinct from cellular necrosis