Human VHL Knockout Cell Line-A549
Cat.No. : CSC-RT2715
Host Cell: A549 Target Gene: VHL
Size: 1x10^6 cells/vial, 1mL Validation: Sequencing
Inquire for Price
Cell Line Information
Cell Culture Information
Safety and Packaging
| Cat. No. | CSC-RT2715 |
| Cell Line Information | This cell is a stable cell line with a homozygous knockout of human VHL using CRISPR/Cas9. |
| Target Gene | VHL |
| Host Cell | A549 |
| Size Form | 1 vial (>10^6 cell/vial) |
| Shipping | Dry ice package |
| Storage | Liquid nirtogen |
| Species | Human |
| 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 |
Inquiry
Background
Case Study
Applications
The Von Hippel-Lindau (VHL) gene, located on chromosome 3p25.3, is essential for suppressing tumor growth. Mutations in this gene cause Von Hippel-Lindau disease, a rare autosomal dominant disorder characterized by the development of multiple benign and malignant tumors in various organ systems. The VHL protein (pVHL) plays a key role in the body's response to oxygen levels. Under normal oxygen conditions, pVHL binds to hypoxia-inducible factor 1-α (HIF1α), leading to its degradation. This process is essential for maintaining cellular homeostasis by regulating genes involved in angiogenesis, erythropoiesis, and glucose metabolism. When pVHL becomes dysfunctional due to genetic mutations, HIF1α accumulates, leading to the transcription of genes that promote tumor growth and angiogenesis.
Clinically, VHL disease presents with a variety of symptoms, depending on the organ system involved. Common manifestations include hemangioblastomas of the central nervous system and retina, renal cell carcinoma, pheochromocytomas, pancreatic cysts, and endolymphatic sac tumors. The variability of symptom presentation requires a high clinical suspicion for prompt diagnosis, especially in individuals with a family history of VHL disease.
The von Hippel-Lindau (VHL) protein binds and degrades hypoxia-inducible factor (HIF), which is hydroxylated by prolyl hydroxylases under normoxic conditions. Although initially described as a tumor suppressor, increasing evidence suggests that VHL may instead promote tumor growth. Here, researchers found that VHL interacts with p53, preventing its tetramerization, promoter binding, and expression of its target genes p21, PUMA, and Bax. VHL limits the decreased proliferation and increased apoptosis caused by p53 activation, independent of prolyl hydroxylation and HIF activity, and its presence in tumors leads to resistance to p53-inducing chemotherapy in vivo. This study suggests that VHL has both antitumor functions through HIF degradation and novel protumor functions through inhibition of p53 targets (p21, PUMA, Bax).
The researchers created two models to study the effects of VHL on p53 activity in vivo: a VHL elimination model (A549 parental cells vs. A549 VHL knockout cells) and a VHL rescue model (−VHL 786-O vs. +VHL 786-O). In each model, VHL-deficient tumor cells were injected into the left flank of nude mice, while VHL-expressing tumor cells were injected into the right flank of the same animals (Figure 1a). These tumors were grown for 4 weeks, their size was measured, and then weekly doxorubicin treatment was initiated for an additional 4 weeks. VHL-deficient tumors initially grew larger than VHL-expressing tumors, likely due to VHL's effects on HIF. However, after induction of p53 with doxorubicin, VHL-deficient tumors responded to treatment to a greater extent than VHL-expressing tumors, as measured by the ratio of post-treatment tumor volume to pre-treatment tumor volume for each animal (Figure 1b, c). Furthermore, p21 protein levels were reduced in VHL-expressing tumors after treatment compared with VHL-deficient tumors, despite similar p53 protein induction in response to doxorubicin (Figure 1d, e). Proliferation was significantly reduced, while apoptosis was significantly increased, in VHL-deficient tumors after treatment. These results suggest that VHL is attenuating the therapeutic effects of anthracyclines on tumors in vivo.
Figure 1. VHL attenuates response to anthracycline chemotherapy in vivo. (Kinnaird A, et al., 2020)
1. Cancer Research: The human VHL knockout cell line (A549) is essential in cancer research, especially in studying renal cell carcinoma and other cancers associated with loss of VHL function. By utilizing this cell line, researchers can study the mechanisms by which VHL mutations lead to tumorigenesis and identify potential therapeutic targets.
2. Hypoxia-Inducible Pathways: VHL is a key regulator of HIF, and knocking out VHL helps understand how cells respond to hypoxic conditions. This research is critical for developing interventions to manipulate the hypoxic response in a variety of diseases, including cancer and ischemic diseases.
3. Drug Screening and Development: By observing how cell lines respond to different compounds, researchers can identify new drugs that may inhibit the growth of VHL-deficient tumors or target other pathways affected by VHL-HIF interactions.
4. Metabolic Studies: VHL proteins are involved in regulating cellular metabolism. Researchers use VHL knockout A549 cells to study metabolic changes that occur due to loss of VHL.
5. Cell Signaling Studies: VHL loss affects several signaling pathways beyond the HIF pathway. By using these cells, scientists can map the complexity of these networks and how they contribute to disease progression and resistance to treatment.
For research use only. Not intended for any clinical use.
