Cat.No. : CSC-RO0897 Host Cell: HepG2
| Cat. No. | CSC-RO0897 |
| Description | This cell line is engineered to stably overexpress human CD274 (also named as PDL1). |
| Gene | CD274 |
| Gene Species | Homo sapiens (Human) |
| Host Cell | HepG2 |
| Host Cell Species | Homo sapiens (Human) |
| Stability | Validated for at least 10 passages |
| Application | 1. Studying the interactions between immune cells and cancer cells 2. Studying the mechanisms of resistance to immune checkpoint blockade 3. High-throughput screening 4. Drug target validation |
| Quality Control | Negative for bacteria, yeast, fungi and mycoplasma. |
| Shipping | Dry ice |
| Storage | Liquid nitrogen |
| Revival | Rapidly thaw cells in a 37°C water bath. Transfer contents into a tube containing pre-warmed media. Centrifuge cells and seed into a 25 cm2 flask containing pre-warmed media. |
| Mycoplasma | Negative |
| Format | One frozen vial containing millions of cells |
| Storage | Liquid nitrogen |
| Safety Considerations |
The following safety precautions should be observed. 1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum. 2. No eating, drinking or smoking while handling the stable line. 3. Wash hands after handling the stable line and before leaving the lab. 4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells. 5. All waste should be considered hazardous. 6. Dispose of all liquid waste after each experiment and treat with bleach. |
| Ship | Dry ice |
Hepatocellular carcinoma (HCC) is the fifth leading cause of cancer-related death worldwide. Programmed cell death ligand 1 (PD-L1 or CD274) is an immune checkpoint protein that binds to programmed cell death ligand 1 (PD-1). PD-1 is expressed in activated T cells and other immune cells and has been used in cancer treatment, including hepatocellular carcinoma (HCC). Recently, PD-L1 has been shown to be overexpressed in drug-resistant cancer cells. Sorafenib is a multikinase inhibitor and the only FDA-approved drug for the treatment of advanced hepatocellular carcinoma. However, some patients show resistance to sorafenib during treatment. This study aimed to evaluate the effect of glucose deprivation on PD-L1 expression in hepatocellular carcinoma cells. The researchers used PD-L1-overexpressing HepG2 cells and IFN-γ-treated SK-Hep1 cells to explore the effect of glycolysis on PD-L1 expression. Results showed that glucose deprivation reduced PD-L1 expression in hepatocellular carcinoma cells. In addition, TCGA data and immunostaining analysis confirmed that the expression of hexokinase II (HK2), which plays a key role in glucose metabolism, was positively correlated with PD-L1. Notably, rapamycin treatment reduced the expression of PD-L1 and HK2 in PD-L1-high-expressing HCC cells and sorafenib-resistant cells. These findings suggest that the regulation of PD-L1 expression by glucose deprivation may be a strategy to overcome PD-L1 upregulation in sorafenib-resistant HCC patients.
The expression levels of PD-L1 were evaluated in four human HCC cell lines after treatment with IFN-γ. After treatment, PD-L1 was significantly upregulated in SK-Hep1 and Hep3B cell lines, but not in Hep2 and Huh7 cell lines. To investigate the effect of glycolysis on PD-L1 expression, PD-L1-overexpressing HepG2 cells (low PD-L1 expression) and IFN-γ-treated SK-Hep1 cells (high PD-L1 expression) were used. The results showed that the expression levels of PD-L1 and HK2 (a marker of glycolysis) were increased in both PD-L1-overexpressing HepG2 cells (Figure 1a, b) and IFN-γ-treated SK-Hep1 cells (Figure 1c, d). In PD-L1-overexpressing HepG2 cells (Figure 1e, f) and IFN-γ-treated SK-Hep1 cells (Figure 1g, h), elevated PD-L1 and HK2 expression levels were reduced after glucose deprivation. These findings suggest a potential association between PD-L1 expression and glycolysis.
Figure 1. Reduced PD-L1 expression after glucose deprivation. (Cho S, et al., 2024)
A: The Human CD274 Stable Cell Line - HepG2 is a specialized cell line derived from the HepG2 hepatocellular carcinoma cell line, stably transfected to express human CD274 (also known as PD-L1). This cell line is typically cultured at 37°C with 5% CO2 in a suitable medium and maintained with appropriate antibiotics in the culture medium. The specific components of the medium and maintenance conditions may vary depending on the laboratory's requirements.
A: The primary application of the Human CD274 Stable Cell Line - HepG2 is to study the association of CD274 (PD-L1) with immune regulation, immune evasion, and immunotherapy. It can be used to investigate CD274 expression, regulatory mechanisms, and its interactions with immune cells, contributing to a better understanding of relevant biological processes in the field of immunotherapy.
A: The CD274 expression in the Human CD274 Stable Cell Line - HepG2 is stable as it is obtained through stable transfection. To verify its stability, methods such as flow cytometry or immunofluorescence staining can be used to detect CD274 expression levels, and periodic checks over multiple passages can confirm that the expression levels remain consistent.
A: Yes, the Human CD274 Stable Cell Line - HepG2 is commonly used for screening and evaluating the efficacy of PD-L1 inhibitors. By using this cell line for drug screening, researchers can assess the effects of candidate drugs on CD274/PD-L1 inhibition, facilitating the discovery of new immunotherapy drugs.
A: To validate the functional activity of CD274 in the Human CD274 Stable Cell Line - HepG2, experimental methods such as ELISA, Western blotting, co-culture with immune cells, and functional assays can be employed to assess CD274 binding to PD-1, signal transduction, and its immunomodulatory effects on immune cells.
A: The Human CD274 Stable Cell Line - HepG2 may be used in immunotherapy vaccine research, especially in the development of vaccines targeting hepatocellular carcinoma. Studying CD274 expression and function in hepatocellular carcinoma cells can help design immune vaccines that enhance the immune system's ability to target HCC.
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