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Real-time quantitative PCR analysis of gene expression in mechanical stimulated invasion assay [extracellular and adhesion]

ABSTRACT: HT1080 human fibrosarcoma cells were seeded onto a 3D collagen-fibronectin matrix and mechanical stimulation in the form of tugging forces was applied to the matrix. Cells not stimulated were used as the control group. The aim of this study was to examine which genes related to ECM and adhesion were deferentially expressed when the mechanical stimulation was applied to the matrix compared to unstimulated, control cells.

ORGANISM(S): Homo sapiens

PROVIDER: GSE98916 | GEO | 2017/05/15

SECONDARY ACCESSION(S): PRJNA388944

REPOSITORIES: GEO

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Project description:The advent of high-throughput measurements of gene expression and bioinformatics analysis methods offers new ways to study gene expression patterns. The primary goal of this study was to determine the time sequence for gene expression in a bone subjected to mechanical loading, during key periods of the bone formation process, including expression of matrix-related genes, the appearance of active osteoblasts, and bone desensitization. A standard model for bone loading was employed in which the right forelimb was loaded axially for three minutes per day, while the left forearm served as a non-loaded, contralateral control. We evaluated loading-induced gene expression over a time course of 4 hours to 32 days after the first loading session. Six distinct, time-dependent patterns of gene expression were identified over the time course and categorized into three primary clusters: genes upregulated early in the time course, genes upregulated during matrix formation, and genes downregulated during matrix formation. Genes were then grouped based on function and/or signaling pathways. Many gene groups known to be important in loading-induced bone formation were identified within the clusters, including AP-1-related genes in the early response cluster, matrix-related genes in the upregulated gene clusters, and Wnt/?-catenin signaling pathway inhibitors in the downregulated gene clusters. Several novel gene groups were identified as well, including chemokine-related genes which were upregulated early but downregulated later in the time course, solute carrier genes which were both up- and downregulated, and muscle-related genes which were primarily downregulated. Time Course with 11 time points, each plus & minus mechanical stimulation with 5 replicates per experimental group (except 12d group which has 4 replicates). Daily mechanical loading was applied to the forearm (24 hours between loading sessions), and ulnae were sampled at indicated time points (4h, 12h, 1d, 2d, 4d, 6d, 8d, 12d, 16d, 24d, or 32d).

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Real-time quantitative PCR analysis of gene expression in mechanical stimulated invasion assay [extracellular and adhesion]

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