• Origin: The Hep G2 cell line is derived from a hepatocellular carcinoma of a 15-year-old, White, male youth with liver cancer. The cell line is commonly used in hepatocellular carcinoma research.
• Background Information: Luciferase-expressing cell lines are genetically engineered to produce luciferase enzymes, typically from fireflies (Photinus pyralis). These bioluminescent reporter systems enable real-time monitoring of cellular processes through light emission when exposed to substrate luciferin. Widely used in molecular biology, they facilitate gene expression studies, drug screening, and in vivo imaging applications. EGFP provides a stable fluorescent marker for reliable visualization and tracking of tumor cells, enabling convenient monitoring of cell growth and tumor progression.
• Gene targeting strategy: The exogenous promoter and luciferase-P2A-EGFP coding sequence were randomly inserted into the Hep G2 cells.
• Tumorigenicity: Confirmed in B-NDG mice.
• Application: The B-Tg(Luc-EGFP) HepG2 tumor model is suitable for preclinical evaluation of antitumor efficacy and therapeutic responses using bioluminescence imaging.

Luminescence signal intensity of B-Tg(Luc-EGFP) Hep G2 cells and EGFP expression analysis in B-Tg(Luc-EGFP) Hep G2 cells. Single cell suspensions from wild-type Hep G2 and B-Tg(Luc-EGFP) Hep G2 #1-B01 cultures were measured using the Bright-GloTM luciferase Assay (Promega, Catalog No. E4030). B-Tg(Luc-EGFP) Hep G2 cells have a strong luminescence signal that is not present in wild-type Hep G2 cells. EGFP was detected on the surface of B-Tg(Luc-EGFP) Hep G2 cells but not wild-type Hep G2 cells.

Subcutaneous tumor growth of B-Tg(Luc-EGFP) Hep G2 cells. B-Tg(Luc-EGFP) Hep G2 cells (1x10⁷) and wild-type Hep G2 cells (1x10⁷) were subcutaneously implanted into B-NDG mice (female, 7-9-week-old, n=6). Tumor volume and body weight were measured twice a week. (A) Average tumor volume. (B) Body weight. Volume was expressed in mm³ using the formula: V=0.5 X long diameter X short diameter². Results indicate that B-Tg(Luc-EGFP) Hep G2 cells were able to establish tumors in vivo and can be used for efficacy studies. Values are expressed as mean ± SEM.

Establishment of orthotopic liver cancer model. To establish the liver cancer model, B-Tg(Luc-EGFP) Hep G2 cell suspension was injected into the left lateral lobe of the liver of B-NDG mice (femal, n=6), and the body weight and tumor fluorescence signal of the mice were recorded weekly. The results showed that the fluorescence signal gradually increased. Right panel showed H&E staining of liver tumors. This indicated that the cell line was successfully constructed as an orthotopic tumor model.

Antitumor activity of anti-HER2 antibody  in B-Tg(Luc-EGFP) Hep G2 liver orthotopic model. The DS-8201 significantly B-Tg(Luc-EGFP) Hep G2 tumor growth in B-NDG mice. B-Tg(Luc-EGFP) Hep G2 cells （5×10⁵）suspension mixed in Matrigel solution (1:1) was orthotopically implanted into B-NDG mice (female, 8 week-old, n=6). Mice were grouped when Imaging signal value reached approximately 2.7×10⁸ p/sec, at which time they were treated with the DS-8201 with different doses and schedules indicated in panel (A) Body weight during treatment. As shown in panel B, this drug was efficacious, demonstrating that B-Tg(Luc-EGFP) Hep G2 liver orthotopic model could provide a powerful preclinical model for in vivo evaluation of DS-8201. Values are expressed as mean ± SEM. **p＜0.01, ****p＜0.0001