Human PTEN Knockout Cell Line-HCT116
Cat.No. : CSC-RT0072
Host Cell: HCT116 Target Gene: PTEN
Size: >1x10^6 cells/vial, 1 mL Validation: Sequencing
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Cell Line Information
Cell Culture Information
Safety and Packaging
| Cat. No. | CSC-RT0072 |
| Cell Line Information | This cell line is a stable cell line with a homozygous knockout of human PTEN using CRISPR/Cas9. |
| Target Gene | PTEN |
| Gene ID | 5728 |
| Genotype | PTEN (-/-) |
| Host Cell | HCT116 |
| Cell Type | Epithelial |
| Size | >1x10^6 cells/vial, 1 mL |
| Sequencing Result | Homozygous: 7 bp deletion in exon |
| Culture Properties | Adherent |
| 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
Applications
The PTEN gene is the most important tumor suppressor gene discovered after the p53 gene. It is located at 10q23.3, with a total length of about 200 kb, including 9 exons and 8 introns, encoding 403 amino acids. Its protein product includes three structural functional regions: an amino-terminal phosphatase region consisting of 180 amino acids, a lipid-binding C2 region, and a carboxyl-terminal region consisting of about 50 amino acids. PTEN gene regulation participates in a variety of cell signaling pathways and can affect tumor growth in many aspects, such as tumor cell proliferation and differentiation, cell cycle, cell apoptosis, and cell migration.
John et al.'s research showed that the inhibitory effect of PTEN on the metastasis, infiltration, and growth of glioma cells is mainly mediated by local focal adhesion kinase (FAK). PTEN regulates FAK in two main ways: first, by dephosphorylating and inactivating FAK, and then downregulating the downstream regulatory signal of FAK, p130CAS, thereby inhibiting cell metastasis, infiltration and growth; second, by inactivating phosphatidylinositol 3-kinase (PI3K), resulting in the inhibition of the PI3K-AKT (protein kinase B) pathway. Masliah-Planchon et al. found that 3-phosphoinositide (PIP3) is an important substrate of PTEN. Functional variation of PTEN protein can lead to the inactivation of lipid phosphatases, but still has protein phosphatase activity. PTEN dephosphorylates PIP3, blocks the PI3K-AKT pathway, reduces the activity of AKT, and accelerates cell apoptosis.
The PTEN gene (phosphatase and tensin homolog) is an important tumor suppressor implicated in a variety of cancers. Knockout (deletion) of this gene in cell lines can aid in understanding the underlying mechanisms of tumorigenesis and the development of targeted therapies. The PTEN knockout HCT116 cell line, derived from human colorectal cancer, is particularly valuable in research. Here are some key applications:
Cancer Research: PTEN loss or mutation is frequently observed in various cancers, making this cell line a valuable model for understanding the molecular mechanisms of cancer progression and resistance to therapies.
Drug Screening: These cells are essential for high-throughput drug screening assays designed to identify new therapeutic compounds. Researchers can evaluate the efficacy of potential drugs targeting pathways altered by PTEN loss, providing insights into new cancer treatment strategies.
Signal Transduction Studies: PTEN is a key regulator of the PI3K/AKT signaling pathway. HCT116 PTEN knockout cells help scientists study the downstream effects of PTEN loss on cellular signaling networks, providing a deeper understanding of its role in cell growth, survival, and metabolism.
Genetic Studies: Using cell lines to evaluate the effectiveness of gene editing tools and techniques in correcting PTEN loss or modulating related pathways. Explore how PTEN deficiency alters the epigenetic landscape of cells and promotes cancer progression.
Biomarker Identification: Researchers use PTEN knockout HCT116 cells to identify and validate biomarkers for cancer diagnosis and prognosis.
Metabolic Studies: The PTEN gene plays a key role in metabolic regulation. Studying PTEN knockout HCT116 cells allows researchers to explore metabolic changes associated with PTEN deficiency, aiding the development of metabolism-based cancer treatments.
For research use only. Not intended for any clinical use.
