Human AKT1/AKT2 Knockout Cell Line-HCT116
Cat.No. : CSC-RT0082
Host Cell: HCT116 Target Gene: AKT1/AKT2
Size: 1x10^6 cells/vial, 1mL Validation: Sequencing
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Cell Line Information
Cell Culture Information
Safety and Packaging
| Cat. No. | CSC-RT0082 |
| Cell Line Information | HCT116 -AKT1/AKT2 (-/-,-/-) is a cell line with a double homozygous knockout of human AKT1 and AKT2 |
| Target Gene | AKT1/AKT2 |
| Host Cell | HCT116 |
| 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 |
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Background
Case Study
Applications
AKT1 and AKT2 are two highly homologous isoforms of the protein kinase B (PKB) family that play crucial roles in various cellular processes, including metabolism, proliferation, cell survival, growth, and angiogenesis. They are encoded by the AKT1 and AKT2 genes, respectively. The two isoforms are highly similar in amino acid sequence and have similar domain structures, including an N-terminal pleckstrin homology (PH) domain, a central kinase domain, and a C-terminal regulatory domain. Despite their similarities, they exhibit different biological functions and regulatory mechanisms. AKT1, also known as PKBα, is ubiquitously expressed in many tissues and is particularly known for its involvement in controlling cell survival and growth. AKT2, also known as PKBβ, has a similar activation mechanism to AKT1 but has a more tissue-specific expression pattern, being primarily expressed in insulin-responsive tissues such as muscle, liver, and adipose tissue. AKT2 plays a key role in regulating glucose homeostasis and insulin signaling.
Dysregulation of the AKT signaling pathway has been implicated in a variety of pathological conditions, including cancer, diabetes, and cardiovascular disease. Overactivation of AKT1 is commonly observed in various cancers, contributing to enhanced cell survival and proliferation. Conversely, mutations or defects in AKT2 are often associated with insulin resistance and type 2 diabetes, underscoring its role in metabolic regulation.
Human tumor growth depends on rapidly dividing cancer cells that drive population expansion. However, even advanced tumors contain slowly proliferating cancer cells for reasons that are unclear. Here, researchers used β1 integrin activation and the AKT1-E17K mutant oncoprotein as in vivo experimental tools to selectively disrupt the ability of rapidly proliferating cancer cells to generate AKT1low daughter cells, which are rare, slowly proliferating, tumor-initiating, and chemoresistant. Surprisingly, researchers found that selective depletion of AKT1low slow proliferators actually reduced growth in a molecularly diverse panel of human cancer cell xenograft models without globally altering cell proliferation or in vivo survival. Furthermore, an unusual cancer patient with an AKT1-E17K mutant solid tumor also failed to generate AKT1low quiescent cancer cells, which was associated with significantly prolonged survival after adjuvant therapy compared to other patients. These findings support a model in which human solid tumor growth depends not only on rapidly proliferating cancer cells but also on the continued generation of AKT1low slow proliferators.
In this study, HCT116-AKT1/2 knockout cells failed to generate quiescent cancer cells (QCCs), but lentiviral-mediated overexpression of cDNA encoding wild-type AKT1 completely restored the generation of these MCM2low/H3K9me2low/HES1high cells in this AKT knockout line (Figure 1). In addition, overexpression of cDNA of myr-AKT1, an artificially mutated kinase-active protein that constitutively localizes to the cell membrane, did not rescue the generation of QCCs in HCT116-AKT1/2 knockout cells compared with wild-type AKT1 (Figure 1).
Figure 1. Bar graph percentages of QCCs in the HCT116-AKT1/2 knockout human cancer cell line with cDNAs for AKT1-WT or AKT1 mutants. (Alves, Cleidson P., et al. 2018)
The creation of the AKT1 and AKT2 knockout HCT116 cell line can be a powerful tool for research in a variety of biomedical fields. Here are some key applications of the human AKT1/AKT2 knockout cell line-HCT116:
Cancer research: This cell line facilitates the study of the role of AKT1 and AKT2 in cancer progression and tumorigenesis, providing insights into mechanisms such as cell survival, proliferation, and metabolic regulation.
Drug screening and development: The knockout model is used for high-throughput screening of AKT inhibitors and other therapeutic agents, helping to identify potential drugs that can selectively target cancer cells with AKT pathway abnormalities.
Signal transduction studies: Researchers use this cell line to dissect signaling pathways regulated by AKT isoforms to better understand how these kinases contribute to cellular function and oncogenic processes.
Metabolic pathway analysis: It is a valuable tool to explore the changes in cellular metabolism that occur upon loss of AKT1/AKT2, revealing metabolic dependencies and vulnerabilities of cancer cells.
Genetic interaction studies: This cell line helps identify genetic interactions and synthetic lethal relationships with AKT pathway components that can be exploited to develop combination therapies for enhanced anticancer efficacy.
Modeling resistance mechanisms: It helps study resistance mechanisms to current therapies and the role of AKT in mediating resistance, with the goal of improving treatment strategies and overcoming therapeutic resistance in cancer treatment.
For research use only. Not intended for any clinical use.
