human osteosarcoma cell line mg 63  (ATCC)

Journal: Molecular Medicine Reports

Article Title: Acetylshikonin induces ferroptosis via the lipid peroxidation pathway in osteosarcoma cells

doi: 10.3892/mmr.2025.13765

Figure Lengend Snippet: Acetylshikonin reduces osteosarcoma cell viability and increases membrane permeability. (A) Molecular structure of acetylshikonin. (B) CCK-8 assay results showing the viability of hFOB 1.19 cells following exposure to acetylshikonin (0.5–3 µM) for 24 h (n=4). (C) MG63, (D) HOS and (E) U2OS cell viability was assessed using the CCK-8 assay following treatment with acetylshikonin (0.05–20 µM) for 24 and 48 h (n=4). (F) Phase-contrast microscopy images depicting morphological changes in osteosarcoma cells treated with acetylshikonin (3 µM) for 24 h (n=4). (G) Fluorescence microscopy images showing nuclear staining with Hoechst 33342, membrane integrity with PI and viability with Calcein-AM in osteosarcoma cells treated with acetylshikonin (0.5–10 µM) for 24 h (n=4). Data are presented as the mean ± SD. *P<0.05 vs. untreated control. CCK-8, Cell Counting Kit-8; PI, propidium iodide.

Article Snippet: The human osteosarcoma cell lines (U2OS, HOS and MG63) and normal human osteoblasts (hFOB 1.19) were sourced from the American Type Culture Collection.

Techniques: Membrane, Permeability, CCK-8 Assay, Microscopy, Fluorescence, Staining, Control, Cell Counting

Journal: Molecular Medicine Reports

Article Title: Acetylshikonin induces ferroptosis via the lipid peroxidation pathway in osteosarcoma cells

doi: 10.3892/mmr.2025.13765

Figure Lengend Snippet: Acetylshikonin induces DNA fragmentation in osteosarcoma cells. Osteosarcoma cells (5×10 5 ) were treated with acetylshikonin (0.1–3 µM) for 24 h, then underwent the TUNEL assay. Fluorescence was analyzed by flow cytometry (n=4). Untreated cells were used as controls. Data are presented as the mean ± SD. *P<0.05, **P<0.01 vs. untreated control.

Article Snippet: The human osteosarcoma cell lines (U2OS, HOS and MG63) and normal human osteoblasts (hFOB 1.19) were sourced from the American Type Culture Collection.

Techniques: TUNEL Assay, Fluorescence, Flow Cytometry, Control

Journal: Molecular Medicine Reports

Article Title: Acetylshikonin induces ferroptosis via the lipid peroxidation pathway in osteosarcoma cells

doi: 10.3892/mmr.2025.13765

Figure Lengend Snippet: Acetylshikonin induces apoptosis in osteosarcoma cells. Osteosarcoma cells (5×10 5 ) were treated with acetylshikonin (0.1–3 µM) for 24 h, then underwent the Annexin V/PI assay (n=4). Untreated cells were used as controls. Data are presented as the mean ± SD. *P<0.05 vs. untreated control. PI, propidium iodide.

Article Snippet: The human osteosarcoma cell lines (U2OS, HOS and MG63) and normal human osteoblasts (hFOB 1.19) were sourced from the American Type Culture Collection.

Techniques: Control

Journal: Molecular Medicine Reports

Article Title: Acetylshikonin induces ferroptosis via the lipid peroxidation pathway in osteosarcoma cells

doi: 10.3892/mmr.2025.13765

Figure Lengend Snippet: Acetylshikonin promotes cell cycle arrest in osteosarcoma cells. Cell cycle distribution of osteosarcoma cells treated with acetylshikonin (0.1–3 µM) for 24 h, was assessed by propidium iodide staining and flow cytometry. Untreated cells were used as controls. Data are presented as the mean ± SD. *P<0.05 vs. untreated control.

Article Snippet: The human osteosarcoma cell lines (U2OS, HOS and MG63) and normal human osteoblasts (hFOB 1.19) were sourced from the American Type Culture Collection.

Techniques: Staining, Flow Cytometry, Control

Journal: Molecular Medicine Reports

Article Title: Acetylshikonin induces ferroptosis via the lipid peroxidation pathway in osteosarcoma cells

doi: 10.3892/mmr.2025.13765

Figure Lengend Snippet: Acetylshikonin promotes intracellular ROS accumulation. Osteosarcoma cells (5×10 5 ) were treated with acetylshikonin (0.1–3 µM) for 1 h, then stained with 1 µM H 2 DCFDA. Fluorescence was analyzed by flow cytometry (n=4). Untreated cells served as controls. Data are presented as the mean ± SD. *P<0.05, **P<0.01 vs. untreated control. ROS, reactive oxygen species.

Article Snippet: The human osteosarcoma cell lines (U2OS, HOS and MG63) and normal human osteoblasts (hFOB 1.19) were sourced from the American Type Culture Collection.

Techniques: Staining, Fluorescence, Flow Cytometry, Control

Journal: Molecular Medicine Reports

Article Title: Acetylshikonin induces ferroptosis via the lipid peroxidation pathway in osteosarcoma cells

doi: 10.3892/mmr.2025.13765

Figure Lengend Snippet: Acetylshikonin disrupts mitochondrial membrane potential. Cells were incubated with acetylshikonin (1 µM) for (A) 2 or (B) 8 h and subsequently stained with JC-1 (n=4). (C) Western blot analysis of Bcl-2, Bcl-xl, Bax and Bak protein expression in osteosarcoma cells following acetylshikonin treatment (0.1–3 µM) for 8 h (n=4). Untreated cells served as controls. Data are presented as the mean ± SD.

Article Snippet: The human osteosarcoma cell lines (U2OS, HOS and MG63) and normal human osteoblasts (hFOB 1.19) were sourced from the American Type Culture Collection.

Techniques: Membrane, Incubation, Staining, Western Blot, Expressing

Journal: Molecular Medicine Reports

Article Title: Acetylshikonin induces ferroptosis via the lipid peroxidation pathway in osteosarcoma cells

doi: 10.3892/mmr.2025.13765

Figure Lengend Snippet: Acetylshikonin decreases mitochondrial volume and enhances lipid peroxidation in osteosarcoma cells. (A) Transmission electron microscopy images of HOS cells treated with acetylshikonin (3 µM) for 24 h, showing a reduction in mitochondrial volume (blue triangles). The red arrow indicates the endoplasmic reticulum. (B) Fluorescence microscopy analysis of lipid peroxidation in osteosarcoma cells treated with acetylshikonin (3 µM) and C11-BODIPY™ 581/591 (n=4). (C-E) Flow cytometric analyses showing lipid peroxidation in osteosarcoma cells incubated with acetylshikonin (0.1–3 µM) and C11-BODIPY (581/591) for 30 min (n=4). (F-H) Intracellular Fe 2+ levels in osteosarcoma cells treated with acetylshikonin (3 µM) for 24 h were quantified using an Fe 2+ detection reagent and microplate reader. (I-K) Western blot analysis of GPX4 protein expression in osteosarcoma cells treated with acetylshikonin (0.1–3 µM) for 8 h (n=4). Untreated cells served as controls. Data are presented as the mean ± SD. *P<0.05 vs. untreated control. GPX4, glutathione peroxidase 4; Fe 2+ , ferrous ion.

Article Snippet: The human osteosarcoma cell lines (U2OS, HOS and MG63) and normal human osteoblasts (hFOB 1.19) were sourced from the American Type Culture Collection.

Techniques: Transmission Assay, Electron Microscopy, Fluorescence, Microscopy, Incubation, Western Blot, Expressing, Control

Journal: Molecular Medicine Reports

Article Title: Acetylshikonin induces ferroptosis via the lipid peroxidation pathway in osteosarcoma cells

doi: 10.3892/mmr.2025.13765

Figure Lengend Snippet: Acetylshikonin induces ferroptosis-mediated cell death. (A-C) CCK-8 assay results of osteosarcoma cells pretreated with ferrostatin-1 (10 µM), z-DEVD-FMK (10 µM), necrostatin-1 (10 µM), IM54 (10 µM) and liproxstain-1 (1 µM) for 1 h before exposure to acetylshikonin (3 µM) for 24 h (n=4). (D) Flow cytometric analysis of Annexin V/PI staining in osteosarcoma cells pretreated with ferrostatin-1 (10 µM) for 1 h followed by acetylshikonin (3 µM) for 24 h (n=4). (E) CCK-8 assay results of osteosarcoma cells treated with acetylshikonin (3 µM), erastin (3 µM) and RSL3 (3 µM) for 24 h (n=4). Untreated cells served as controls. Data are presented as the mean ± SD. *P<0.05 vs. untreated control; # P<0.05 vs. acetylshikonin-treated group. CCK-8, Cell Counting Kit-8.

Article Snippet: The human osteosarcoma cell lines (U2OS, HOS and MG63) and normal human osteoblasts (hFOB 1.19) were sourced from the American Type Culture Collection.

Techniques: CCK-8 Assay, Staining, Control, Cell Counting