Journal: Cancer Management and Research

Article Title: Regorafenib-Induced Stress Response Alters the Bioenergetic Profile of Osteosarcoma Cells and Modulates Gene Expression Associated with Metabolic regulation-a Potential Mechanism of Osteosarcoma Treatment-Related Adaptation

doi: 10.2147/CMAR.S562346

Figure Lengend Snippet: Representative dose‒response curves used to determine the IC50 values for a well-established osteosarcoma cell line, MG63 (REG IC 50 26±3 µM) ( a ), and primary cells, APR1 (REG IC 50 42±12 µM) ( b ). Data are presented as mean ± SD from n = 3 biological replicates, each performed in 4 technical replicates.

Article Snippet: Regorafenib activity was analyzed in vitro using a well-established human osteosarcoma cell line, MG63 (ATCC; CRL-1427), and primary APR1 cells established from patient tissue.

Techniques:

Journal: Cancer Management and Research

Article Title: Regorafenib-Induced Stress Response Alters the Bioenergetic Profile of Osteosarcoma Cells and Modulates Gene Expression Associated with Metabolic regulation-a Potential Mechanism of Osteosarcoma Treatment-Related Adaptation

doi: 10.2147/CMAR.S562346

Figure Lengend Snippet: Visualization of the morphology of MG63 and APR1 osteosarcoma cells by epifluorescence microscopy under control (DMSO-treated) and regorafenib-treated (IC50) conditions. Key cellular structures were visualized via fluorescent dyes: the nuclei were stained with DAPI (blue), the actin cytoskeleton was stained with phalloidin Atto-488 (green), and the mitochondrial network was stained with mitoRed (red). Magnification 100x, scale bar: 200 µm; 200x, scale bar: 100 µm.

Article Snippet: Regorafenib activity was analyzed in vitro using a well-established human osteosarcoma cell line, MG63 (ATCC; CRL-1427), and primary APR1 cells established from patient tissue.

Techniques: Epifluorescence Microscopy, Control, Staining

Journal: Cancer Management and Research

Article Title: Regorafenib-Induced Stress Response Alters the Bioenergetic Profile of Osteosarcoma Cells and Modulates Gene Expression Associated with Metabolic regulation-a Potential Mechanism of Osteosarcoma Treatment-Related Adaptation

doi: 10.2147/CMAR.S562346

Figure Lengend Snippet: The invasion of the osteosarcoma cell lines MG63 and APR1 is modulated by regorafenib. Representative images ( a ) and quantitative analysis ( b ) illustrating the invasive capacity of MG63 and APR1 cells following treatment with regorafenib at the IC50 concentration. Magnification: 40-fold and 100-fold; scale bar: 100 µm. Statistical significance was indicated via asterisks ****p<0.0001. Data are shown as mean ± SD based on three independent biological replicates, each assessed in two technical replicates, with two images per replicate subjected to ImageJ analysis. Results are expressed as the percentage of migrated cells per field and normalized to the corresponding control conditions.

Article Snippet: Regorafenib activity was analyzed in vitro using a well-established human osteosarcoma cell line, MG63 (ATCC; CRL-1427), and primary APR1 cells established from patient tissue.

Techniques: Concentration Assay, Control

Journal: Cancer Management and Research

Article Title: Regorafenib-Induced Stress Response Alters the Bioenergetic Profile of Osteosarcoma Cells and Modulates Gene Expression Associated with Metabolic regulation-a Potential Mechanism of Osteosarcoma Treatment-Related Adaptation

doi: 10.2147/CMAR.S562346

Figure Lengend Snippet: The DNA content distribution in the osteosarcoma cell lines was assessed via flow cytometry. Representative histograms ( a ) illustrate the proliferative status of cells across the cell cycle phases in the control and regorafenib-treated groups. The cells were classified into three distinct populations: G0/G1 phase, S phase, and G2/M phase ( b ). Statistically significant differences are indicated by asterisks (**p < 0.01 and ***p < 0.001), whereas comparisons with no statistically significant differences are marked as “ns”. Values represent the mean ± SD derived from two independent biological replicates, each including three technical replicates.

Article Snippet: Regorafenib activity was analyzed in vitro using a well-established human osteosarcoma cell line, MG63 (ATCC; CRL-1427), and primary APR1 cells established from patient tissue.

Techniques: Flow Cytometry, Control, Derivative Assay

Journal: Cancer Management and Research

Article Title: Regorafenib-Induced Stress Response Alters the Bioenergetic Profile of Osteosarcoma Cells and Modulates Gene Expression Associated with Metabolic regulation-a Potential Mechanism of Osteosarcoma Treatment-Related Adaptation

doi: 10.2147/CMAR.S562346

Figure Lengend Snippet: Effect of regorafenib at the IC 5 0 concentration on the expression profile of genes associated with cancer-related pathways in MG63 ( a ) and APR1 ( b ) osteosarcoma cell lines. The x-axis of volcano plot represents log 2 (fold regulation) and the y-axis −log 1 0 (p-value). Upregulated genes are marked with red upward arrows, whereas downregulated genes are marked with green downward arrows. Genes consistently up- or downregulated in both MG-63 and APR-1 cells are additionally highlighted with light-red or light-green circles, respectively. Vertical dashed lines indicate the fold-change cutoff used to define differential expression.Genes showing expression changes of at least 2-fold were classified as differentially expressed. Gene expression levels were normalized to those of reference genes ( ACTB , B2M , RPLP0 , and GAPDH ) and calculated using the 2^(-ΔΔCt) method. The reactions for this assay were performed using three independent biological replicates, each processed as one technical replicate.

Article Snippet: Regorafenib activity was analyzed in vitro using a well-established human osteosarcoma cell line, MG63 (ATCC; CRL-1427), and primary APR1 cells established from patient tissue.

Techniques: Concentration Assay, Expressing, Quantitative Proteomics, Gene Expression

Journal: Cancer Management and Research

Article Title: Regorafenib-Induced Stress Response Alters the Bioenergetic Profile of Osteosarcoma Cells and Modulates Gene Expression Associated with Metabolic regulation-a Potential Mechanism of Osteosarcoma Treatment-Related Adaptation

doi: 10.2147/CMAR.S562346

Figure Lengend Snippet: Heatmap illustrating the expression profiles of all 84 genes involved in pathways associated with oncogenesis. The heatmap provides a comprehensive overview of gene expression modulation in MG63 and APR1 osteosarcoma cell lines following treatment with regorafenib at the IC50 concentration.

Article Snippet: Regorafenib activity was analyzed in vitro using a well-established human osteosarcoma cell line, MG63 (ATCC; CRL-1427), and primary APR1 cells established from patient tissue.

Techniques: Expressing, Gene Expression, Concentration Assay