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Conditioned medium of PC-3 prostate cancer influences mRNA profiles in mechanically strained osteoblasts

ABSTRACT: Bone is the main site of metastasis from prostate cancer, it is important to investigated miRNAs and mRNAs of bone metastases from prostate cancer. Considering that bone is in an appropriate mechanical environment in physiological state, in this study, the miRNA, mRNA, lncRNA profiles of mechanically strained osteoblasts treated with conditioned medium of PC-3 prostate cancer cells were studied. MC3T3-E1 osteoblastic cells were treated with conditioned medium of PC-3 prostate cancer cells, at the same time stimulated with mechanical tensile strain of 2,500 microstrain (με) at 0.5 Hz, the osteoblastic differentiation of the MC3T3-E1 cells were assayed

ORGANISM(S): Mus musculus

PROVIDER: GSE211413 | GEO | 2022/08/23

REPOSITORIES: GEO

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Project description:Bone defects are a global public health problem. However, the available methods for bone regeneration are limited. In recent years, the application of traditional Chinese herbs for bone regeneration has attracted increasing attention. β-ecdysterone is a herbal extract similar to estrogen that promotes the metabolism of cell protein synthesis; however, its function on bone regeneration is not yet clear. In this study, we investigated the function of β-ecdysterone on osteoblast differentiation and bone regeneration in vitro and in vivo. MC3T3-E1 cells were used to test the function of β-ecdysterone on osteoblast differentiation and bone regeneration in vitro. The results of the CCK-8 assay suggested that the proliferation of MC3T3-E1 cells was promoted by β-ecdysterone. Furthermore, β-ecdysterone influenced the expression of osteogenesis-related genes, and the bone regeneration capacity of MC3T3-E1 cells was detected by PCR, the alkaline phosphatase (ALP) test, and the Alizarin Red test. β-ecdysterone could up-regulate the expression of osteoblastic-related genes, and promoted ALP activity and the formation of calcium nodules. Moreover, we also determined that β-ecdysterone increased the mRNA and protein levels of components of the BMP-2/Smad/Runx2/Osterix pathway. DNA sequencing further confirmed these target effects. β-ecdysterone promoted bone formation by enhancing gene expression of the BMP-2/Smad/Runx2/Osterix signaling pathway and by the enrichment biological processes. For in vivo experiments, the femur condyle defection model was constructed by drilling a critical-sized (3×3 mm) defect in 12-week-old SD male rats to further assess the bone regeneration functions of β-ecdysterone. The results of micro-CT showed that β-ecdysterone could accelerate bone regeneration and exhibited higher bone volume, bone surface, bone mineral density at each observation time point. Immunohistochemistry confirmed that the β-ecdysterone also increased the expression of collagen, osteocalcin, and BMP2 in the experiment group at 8 weeks. In conclusion, discovered β-ecdysterone is a new bone regeneration regulator that can stimulate MC3T3-E1 cell proliferation and induce bone regeneration through the BMP-2/Smad/Runx2/Osterix pathway. This new function of β-ecdysterone has revealed a new direction of osteogenic differentiation and has provided novel therapeutic strategies for treating bone defection.

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Conditioned medium of PC-3 prostate cancer influences mRNA profiles in mechanically strained osteoblasts

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