HMGB1 Knockout Cell Line-HeLa
Cat.No. : CSC-RT1762
Host Cell: HeLa Target Gene: HMGB1
Size: 1x10^6 cells/vial, 1mL Validation: Sequencing
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Cell Line Information
Cell Culture Information
Safety and Packaging
| Cat. No. | CSC-RT1762 |
| Cell Line Information | HeLa -HMGB1(-/-) is a stable cell line with a homozygous knockout of human HMGB1 using CRISPR/Cas9. |
| Target Gene | HMGB1 |
| Host Cell | HeLa |
| Shipping | 1 vial of knockout cell line |
| Storage | Liquid nitrogen |
| Species | Human |
| Gene Symbol | HMGB1 |
| Gene ID | 3146 |
| 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. |
| Media Type | Cells were cultured in DMEM supplemented with 10% fetal bovine serum. |
| Growth Properties | Cells are cultured as a monolayer at 37°C in a humidified atmosphere with 5% CO2. Split at 80-90% confluence, approximately 1:4-1:6. |
| Freeze Medium | Complete medium supplemented with 10% (v/v) DMSO |
| 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 |
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Background
Case Study
Applications
High mobility group box 1 (HMGB1) is a highly conserved non-histone nuclear protein that plays a crucial role in various biological processes, including regulation of gene expression, DNA repair, and cellular responses to stress. HMGB1 is ubiquitously expressed in almost all cell types and consists of 215 amino acids, including two DNA binding domains (A and B boxes) and an acidic tail. Within the nucleus, HMGB1 acts as a structural chromatin-binding protein that promotes DNA bending, thereby facilitating the assembly of nucleoprotein complexes essential for transcription, replication, and repair. By binding to distorted DNA structures, such as those found at sites of damage, HMGB1 can stabilize nucleosomes and regulate the activity of various transcription factors.
In addition to its nuclear functions, HMGB1 can be actively secreted by immune cells or passively released by necrotic cells during periods of cellular stress or damage. In the extracellular environment, HMGB1 is a potent pro-inflammatory cytokine. It interacts with various receptors, including the receptor for advanced glycation end products (RAGE) and Toll-like receptors (TLR2 and TLR4), to initiate and propagate inflammatory responses. This extracellular action of HMGB1 is critical in infection, injury, and immune responses, and it contributes to the pathogenesis of various inflammatory and autoimmune diseases.
Tumor recurrence during treatment is an important cause of treatment failure. Here, researchers revealed a new mechanism by which high-mobility group box 1 (HMGB1) released by dying cells during radiotherapy or chemotherapy can stimulate the proliferation of viable tumor cells. Inhibition or genetic ablation of HMGB1 inhibited tumor cell proliferation. This effect was due to HMGB1 binding to members of the receptor for advanced glycation end products (RAGE), thereby activating downstream ERK and p38 signaling pathways and promoting cell proliferation. In addition, higher levels of HMGB1 expression in tumor tissues were associated with poor overall survival, and higher concentrations of HMGB1 were detected in the serum of patients who received radiotherapy. Together, these findings suggest that the interaction between dying and surviving cells may affect the fate of tumors. HMGB1 may be a novel tumor promoter with therapeutic and prognostic implications for cancer.
In this study, HT29 and HeLa cells showed elevated levels of ERK signaling activation after radiation, which became less obvious in HMGB1 knockout tumor cells (Figure 1a, b). Similar induction was also observed in HMGB1 WT tumor cells treated with culture supernatants of irradiated HMGB1 WT or HMGB1 knockout cells, although the activation time and intensity were slightly different (Figure 1a, b). γH2AX is a well-known marker of DNA damage. In this study, irradiated tumor cells showed different degrees of γH2AX protein expression, while HMGB1 knockout tumor cells showed severe DNA damage (Figure 1a, b). In addition, radiation did not induce JNK activation in both HeLa and HT29 cells, while p38 showed a short-term activation. (Figure 1a, b). Overall, the downstream molecules of HMGB1/RAGE are mainly involved in the ERK and p38 signaling pathways, while the JNK signaling pathway may not be involved.
Figure 1. Extracellular HMGB1 activates ERK and p38 signaling pathway to promote surviving cell proliferation. a Left panel, expression changes of ERK, JNK, p38, and γH2AX in irradiated HT29 HMGB1 WT and HT29 HMGB1 KO cells. Right panel, expression changes of ERK, JNK, p38 and γ-H2AX in HeLa HMGB1 WT and HeLa HMGB1 KO cells. b Left panel, expression changes of ERK, JNK and p38 in HT29 HMGB1 WT cells treated by 10Gy-irradiated HT29 HMGB1 WT or HT29 HMGB1 KO cell culture supernatant collected from indicated time points. Right panel, expression changes of ERK, JNK and p38 in HeLa HMGB1 WT cells treated by 10Gy-irradiated HeLa HMGB1 WT or HeLa HMGB1 KO cell culture supernatant collected from indicated time points. (He S, et al. 2018)
High Mobility Group Box 1 (HMGB1) is a non-histone chromatin-binding protein that plays significant roles in DNA architecture and various cellular processes such as transcription, DNA repair, and responses to stress and damage. HMGB1 knockout HeLa cell lines can be utilized in numerous research applications:
Cancer Research: HMGB1 is known to be involved in tumor growth and metastasis. By utilizing an HMGB1 knockout HeLa cell line, researchers can study its specific role in cancer development and progression.
Inflammation Studies: HMGB1 plays a crucial role in inflammation and the immune response. Knockout HeLa cell lines enable detailed investigation into how the absence of HMGB1 affects inflammatory pathways, which is valuable for developing anti-inflammatory drugs and treatments for inflammatory diseases.
Cell Death Mechanisms: HMGB1 is involved in various forms of cell death, including apoptosis, necrosis, and autophagy. Using an HMGB1 knockout HeLa cell line allows researchers to dissect the pathways and molecular interactions involved in these processes, contributing to the understanding of cell death mechanisms.
Drug Screening: An HMGB1 knockout HeLa cell line can be used to screen potential therapeutic compounds that target pathways involving HMGB1. This application is particularly relevant for identifying drugs that could modulate HMGB1's effects in diseases such as cancer and autoimmune disorders.
DNA Repair Research: Studying HMGB1 knockout HeLa cells provides insights into how the loss of HMGB1 impacts DNA damage repair and genomic stability.
For research use only. Not intended for any clinical use.
