human hcc cell lines hepg2  (ATCC)

Journal: Journal of Translational Medicine

Article Title: The Achilles’ heel of hepatocellular carcinoma: ginsenoside compound K as a novel GPX4 degrader promotes ferroptosis in hepatocellular carcinoma

doi: 10.1186/s12967-025-07587-9

Figure Lengend Snippet: CK inhibits HCC cells proliferation in vitro. ( A - B ) Hepatoma cell lines (HepG2 and Hep3B) were treated with varying concentrations of CK for 24 h, and cell viability was assessed using the CCK8 assay. ( C - E ) HepG2 cells were cultured with 5µM, 10µM, or 20µM CK for 24 h, followed by measurement of intracellular GSH, MDA, and Fe²⁺ levels. ( F - H ) Similarly, Hep3B cells were cultured with 5µM, 10µM, or 20µM CK for 24 h, and intracellular GSH, MDA, and Fe²⁺ levels were determined. ( I - J ) Both HepG2 and Hep3B cells were cultured with 5µM, 10µM, or 20µM CK for 24 h, and intracellular ROS levels were measured. Data are means ± S .D. and are representative of three independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: All cell lines used in this study, including the human embryonic kidney cell line HEK293T and the human hepatocellular carcinoma (HCC) cell lines HepG2 and Hep3B, were purchased from the American Type Culture Collection (ATCC).

Techniques: In Vitro, CCK-8 Assay, Cell Culture

Journal: Journal of Translational Medicine

Article Title: The Achilles’ heel of hepatocellular carcinoma: ginsenoside compound K as a novel GPX4 degrader promotes ferroptosis in hepatocellular carcinoma

doi: 10.1186/s12967-025-07587-9

Figure Lengend Snippet: CK Induces Ferroptosis in HCC Cells. ( A - B ) Cell viability of HepG2 and Hep3B cells exposed to 10µM CK, with or without Fer-1 (10µM), Lip-1 (1µM), DFO (100µM), DFX (20µM), NAC (40µM), or Trolox (20µM) for 24 h, was assessed using the CCK8 assay. ( C ) Cell viability of hepatoma cells treated with 10µM CK (with or without NAC and Trolox) for 24 h was measured by CCK8. ( D - E ) Intracellular GSH and MDA levels were determined in hepatoma cells treated with 10µM CK (with or without NAC and Trolox) for 24 h. ( F ) Cysteine uptake capacity was evaluated in HepG2 cells treated with 10µM CK for 24 h. ( G ) Cell viability of HepG2 cells treated with various concentrations of CK (with or without cysteine) for 24 h was assessed by CCK8. ( H ) Intracellular GSH content was measured in HepG2 cells treated with 10µM CK (with or without cysteine) for 24 h. ( I ) Mitochondrial morphology and structure of HepG2 cells treated with 10µM CK, 10µM CK + Fer-1 (10µM), or Erastin (20µM) for 24 h were observed using transmission electron microscopy. Data are means ± S.D. and are representative of three independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: All cell lines used in this study, including the human embryonic kidney cell line HEK293T and the human hepatocellular carcinoma (HCC) cell lines HepG2 and Hep3B, were purchased from the American Type Culture Collection (ATCC).

Techniques: CCK-8 Assay, Transmission Assay, Electron Microscopy

Journal: Journal of Translational Medicine

Article Title: The Achilles’ heel of hepatocellular carcinoma: ginsenoside compound K as a novel GPX4 degrader promotes ferroptosis in hepatocellular carcinoma

doi: 10.1186/s12967-025-07587-9

Figure Lengend Snippet: CK promotes ferroptosis by inhibiting GPX4 enzymatic activity. ( A - B ) Protein expression levels of GPX4, SOD, CAT, and TXN in HepG2 and Hep3B cells were detected by Western blotting (WB) after exposure to 5µM, 10µM, or 20µM CK for 24 h. ( C - D ) Semi-quantitative analysis of the protein expression levels mentioned above. ( E - F ) The mRNA levels of GPX4 in HepG2 and Hep3B cells were measured by quantitative real-time PCR (qPCR) after treatment with 5µM, 10µM, or 20µM CK for 24 h. ( G - H ) Localization and expression of GPX4 in HepG2 and Hep3B cells were detected by immunofluorescence (IF) following exposure to 5µM, 10µM, or 20µM CK for 24 h. ( I - J ) Semi-quantitative analysis of the immunofluorescence results mentioned above. ( K - N ) Relative levels of lipid peroxides in HepG2 and Hep3B cells treated with low- and high-dose CK, Sorafenib, or Fer-1. Data are means ± S.D. and are representative of three independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: All cell lines used in this study, including the human embryonic kidney cell line HEK293T and the human hepatocellular carcinoma (HCC) cell lines HepG2 and Hep3B, were purchased from the American Type Culture Collection (ATCC).

Techniques: Activity Assay, Expressing, Western Blot, Real-time Polymerase Chain Reaction, Immunofluorescence

Journal: Journal of Translational Medicine

Article Title: The Achilles’ heel of hepatocellular carcinoma: ginsenoside compound K as a novel GPX4 degrader promotes ferroptosis in hepatocellular carcinoma

doi: 10.1186/s12967-025-07587-9

Figure Lengend Snippet: Overexpression of GPX4 rescues the ferroptosis-promoting effect of CK in HCC cells. ( A - B ) WB analysis of GPX4 protein expression in HepG2 and Hep3B cells after transfection with an overexpression plasmid for GPX4, along with its semi-quantitative analysis. ( C ) PCR detection of GPX4 mRNA levels in cells transfected with the GPX4 overexpression plasmid. ( D - E ) Cell viability of HCC cells transfected with either the GPX4 overexpression plasmid or a vector control, and exposed or not exposed to 10µM CK for 24 h. ( F - G ) GSH content in HCC cells transfected with the GPX4 overexpression plasmid or vector control, and exposed or not exposed to 10µM CK for 24 h. ( H - I ) ROS levels in HCC cells transfected with the GPX4 overexpression plasmid or vector control, and exposed or not exposed to 10µM CK for 24 h. Data are means ± S.D. and are representative of three independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: All cell lines used in this study, including the human embryonic kidney cell line HEK293T and the human hepatocellular carcinoma (HCC) cell lines HepG2 and Hep3B, were purchased from the American Type Culture Collection (ATCC).

Techniques: Over Expression, Expressing, Transfection, Plasmid Preparation, Control

Journal: Journal of Translational Medicine

Article Title: The Achilles’ heel of hepatocellular carcinoma: ginsenoside compound K as a novel GPX4 degrader promotes ferroptosis in hepatocellular carcinoma

doi: 10.1186/s12967-025-07587-9

Figure Lengend Snippet: CK modulates glutamine metabolism in tumor tissues to trigger Ferroptosis. ( A - B ) Intracellular deubiquitinase activity was assayed in HepG2 and Hep3B cells after 24-hour treatment with various doses of CK and the deubiquitinates inhibitor NEM (20µM). ( C ) Principal component analysis (PCA) plot of transcriptome sequencing data from Ctrl and CK groups ( n = 3). ( D ) Volcano plot showing differentially expressed genes between Ctrl and CK groups. ( E ) Heatmap of the top 20 differentially expressed genes. ( F ) Venn diagram illustrates the intersection between differentially expressed genes and a deubiquitinase database. ( G ) Bar chart of Gene Ontology (GO) enrichment analysis for differentially expressed genes. ( H ) Chord diagram of Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis for differentially expressed genes. ( I ) Gene Set Enrichment Analysis (GSEA) plot for differentially expressed genes

Article Snippet: All cell lines used in this study, including the human embryonic kidney cell line HEK293T and the human hepatocellular carcinoma (HCC) cell lines HepG2 and Hep3B, were purchased from the American Type Culture Collection (ATCC).

Techniques: Activity Assay, Sequencing

Journal: Translational Oncology

Article Title: β-Sitosterol enhances the anti-tumor efficacy of sorafenib in hepatocellular carcinoma via the FXR/LXR/ SREBP1/ FASN pathway

doi: 10.1016/j.tranon.2025.102610

Figure Lengend Snippet: Antitumor effects of SIT and combined treatment on HCC cells. HepG2 cells (A) and Hepa1–6 cells (B) were administrated SIT, sorafenib, and sorafenib + SIT at serial concentrations, and Cell Counting Kit-8 (CCK-8) assays were used to determine the cell viability. HepG2 cells (C) and Hepa1–6 cells (D) were treated with SIT (2 μM), sorafenib (10 μM) or a combination of both, and cell proliferation was evaluated using the 5-ethynyl-29-deoxyuridine (EdU) assay. HepG2 cells (E) and Hepa1–6 cells (F) were treated with SIT (2 μM), sorafenib (10 μM) or a combination of both, and cell apoptosis induction was evaluated using flow cytometry. Hepa1–6 cells (G) were treated with SIT (2 μM), sorafenib (10 μM) or a combination of both, and vasculogenic mimicry (VM) formation was evaluated in three-dimensional culture. Data were presented as mean ± SEM, n = 3, One-way ANOVA. * p < .05; ** p < .01; *** p < .001; and **** p < .0001.

Article Snippet: The human HCC cell line HepG2 and the murine HCC cell line Hepa1–6 was purchased from the American Type Culture Collection.

Techniques: Cell Counting, CCK-8 Assay, EdU Assay, Flow Cytometry