Human EGFR-H773_V774insNPH Stable Cell Line-Ba/F3

Name: Human EGFR-H773_V774insNPH Stable Cell Line-Ba/F3
SKU: CSC-RO0135
Availability: InStock

For research use only. Not intended for any clinical use.

Cat. No. : CSC-RO0135

Host Cell : Ba/F3 Size : >1x10⁶ frozen cells/vial

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Cell Line Information

Cell Culture Information

Safety and Packaging

Gene Information

Cat. No.	CSC-RO0135
Description	Ba/F3-EGFR-H773_V774insNPH cell line is a stably transfected cell line which expresses human epidermal growth factor receptor (EGFR) with H773_V774insNPH mutation.
Target Gene	EGFR
Gene Species	Homo sapiens (Human)
Host Cell	Ba/F3
Host Cell Species	Mus musculus (Mouse)
Applications	Drug screening and biological assays
Biosafety Level	2
Size	>1x10^6 frozen cells/vial, 1 mL
Stability	Stable in culture over a minimum of 10 passages
Quality Control	Negative for bacteria, yeast, fungi and mycoplasma.
Growth Conditions	37 °C, 5% CO₂
Shipping	Dry ice

Thawing & Subculturing Instructions	1. Thaw cells by gently swirling in a 37°C water bath. To limit contamination, do not submerge the O-ring and cap. 2. When cells are ~70% thawed (~1 min), transfer the vial into a biosafety cabinet, and wipe the surface with 70% ethanol. Allow tube to dry completely. 3. Transfer the cells gently into a 15 mL conical tube containing 10 mL of pre-warmed culture medium (without antibiotic selection marker). Centrifuge cells at ~125 x g for 5~7 min. 4. Remove supernatant without disturbing the pellet, and resuspend cells in 1 mL culture medium (without antibiotic selection marker). Transfer cells to a 6-well plate containing ~2 mL pre-warmed growth medium (without antibiotic selection marker) or a T25 flask containing 5 mL pre-warmed culture medium (without antibiotic selection marker). 5. Incubate the culture at 37°C with 5% CO2. 6. Subculture: split saturated culture 1:4 ~ 1:6 every 3 days; seed out at about 1~3 x 10^5 cells/mL.
Freeze Medium	Frozen with 70% medium, 20% FBS, 10% DMSO
Growth Properties	Suspension, round
Freezing Instructions	Cells are recommended to generate additional frozen stocks at early passages. Frozen stocks should be preserved in a designated cryopreservation medium or in 70% RPMI 1640 + 20% FBS + 10% DMSO (without antibiotic selection marker). 1. Prepare the freezing medium (70% RPMI 1640 + 20% FBS + 10% DMSO, without antibiotic selection marker) fresh immediately before use. 2. Keep the freezing medium on ice and label cryovials. 3. Transfer cells to a sterile, conical centrifuge tube, and count the cells. 4. Centrifuge the cells at 250 x g for 5 minutes at room temperature and carefully aspirate off the medium. 5. Resuspend the cells at a density of at least 3 x10^6 cells/ml in chilled freezing medium. 6. Aliquot 1 ml of the cell suspension into each cryovial. 7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer. 8. Transfer vials to liquid nitrogen for long-term storage.

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

Gene Name	EGFR epidermal growth factor receptor [ Homo sapiens ]
Gene Symbol	EGFR
Synonyms	ERBB; HER1; mENA; ERBB1; PIG61
Gene Description	epidermal growth factor receptor (erythroblastic leukemia viral (v-erb-b) oncogene homolog, avian)
Gene ID	1956
Uni Prot ID	P00533
m RNA Refseq	NM_005228.3
Protein Refseq	NP_005219.2
Chromosome Location	7p12
Function	ATP binding; MAP kinase kinase kinase activity; actin filament binding; double-stranded DNA binding; enzyme binding; epidermal growth factor-activated receptor activity; epidermal growth factor-activated receptor activity; identical protein binding; contributes_to nitric-oxide synthase regulator activity; protein binding; protein heterodimerization activity; protein phosphatase binding; protein tyrosine kinase activity; protein tyrosine kinase activity; protein tyrosine kinase activity; receptor signaling protein tyrosine kinase activity; transmembrane receptor protein tyrosine kinase activity; transmembrane signaling receptor activity;
Pathway	Adaptive Immune System, organism-specific biosystem; Adherens junction, organism-specific biosystem; Adherens junction, conserved biosystem; Alpha6-Beta4 Integrin Signaling Pathway, organism-specific biosystem; Androgen Receptor Signaling Pathway, organism-specific biosystem; Arf6 signaling events, organism-specific biosystem; Axon guidance, organism-specific biosystem;
MIM	131550

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Background

Case Study

Applications

Q & A

Customer Reviews

The EGFR-H773_V774insNPH gene mutation is a specific alteration that occurs in the Epidermal Growth Factor Receptor (EGFR) gene, which is responsible for coding the EGFR protein. This protein is a type of receptor tyrosine kinase that plays a critical role in cell growth, proliferation, and differentiation in response to the binding of epidermal growth factors. Mutations in the EGFR gene can lead to uncontrolled cell growth and are commonly found in various cancers, such as lung, colon, and breast cancers.

The EGFR-H773_V774insNPH mutation is a unique insertion mutation that occurs between amino acids 773 and 774 in the EGFR protein sequence. This mutation results in the insertion of the sequence “NPH” (asparagine-proline-histidine) within the protein. This alteration disrupts the normal structure and function of the EGFR protein, leading to its constitutive activation. Constitutive activation of the EGFR protein stimulates continuous signaling pathways, promoting cell growth, survival, and migration.

This specific mutation is often associated with acquired resistance to EGFR inhibitors, such as gefitinib and erlotinib, which are commonly used in the treatment of EGFR-mutant lung cancers. The EGFR-H773_V774insNPH mutation is particularly concerning as it confers a poor prognosis and limited treatment options for patients.

In non-small-cell lung cancer (NSCLC), uncommon epidermal growth factor receptor (EGFR) mutations account for 10%–20% of all EGFR mutations. Mutations often impair the efficacy of traditional medicines like osimertinib and afatinib. Thus, novel EGFR-tyrosine kinase inhibitors are needed. Aumolertinib, a third-generation EGFR-TKI, may cure advanced NSCLC, however its effectiveness against uncommon mutations is uncertain. In this work, engineered Ba/F3 cells and patient-derived cells with diverse rare EGFR mutations were used to explore in vitro anticancer efficacy of aumolertinib. Results indicated that aumolertinib effectively inhibited the viability of cell lines harboring these mutations, outperforming its effects on wild-type EGFR cells. Additionally, aumolertinib demonstrated significant tumor growth inhibition in mouse models with specific uncommon mutations, suggesting its potential as a viable treatment option.

Figure 1 illustrates the antitumor effects of aumolertinib in a patient-derived xenograft (PDX) model with the EGFR H773-V774insNPH mutation, showcasing tumor volume, weight, body weights of mice, tumor photographs, and protein expression changes in response to treatment. (doi: 10.1016/j.apsb.2023.03.007) Figure 1. The researchers evaluated the antitumor activity of aumolertinib using a patient-derived xenograft (PDX) model featuring the EGFR H773-V774insNPH mutation. (Shi C, et al., 2016)

Creative Biogene's Human EGFR-H773_V774insNPH Stable Cell Line - Ba/F3 provides a valuable tool for further exploring the therapeutic implications of aumolertinib and similar agents. Our cell line may facilitate rigorous testing of new compounds, ensuring that emerging therapies can be evaluated for effectiveness against hard-to-treat NSCLC variants.

Epidermal Growth Factor Receptor (EGFR)-H773_V774 gene mutation is a specific alteration in the Epidermal Growth Factor Receptor (EGFR) gene, which is involved in cell growth, survival and fragmentation. This mutation involves the conversion of histidine to valine at positions 773 and 774 in the epidermal growth factor receptor protein, which can lead to constitutive activation of the receptor, promote uncontrolled cell growth and lead to the development of cancer.

The epidermal growth factor receptor-H773_V774 mutation is particularly common in certain types of lung cancer (e.g., adenocarcinoma) and is often associated with a poor prognosis. Targeting this mutation has become an important strategy for the treatment of epidermal growth factor receptor-positive lung cancer.

Targeted therapies such as tyrosine kinase inhibitors (TKIs) have been developed to block the activity of mutated epidermal growth factor receptor proteins. These inhibitors, such as ositinib, dacomitinib, and gefitinib, have shown efficacy in treating epidermal growth factor receptor-H773_V774 mutant lung cancers, improving patient prognosis and prolonging survival.

In addition to its role in cancer treatment, the epidermal growth factor receptor-H773_V774 mutation can be used as a predictive biomarker to guide treatment choices for lung cancer patients. Detection of this mutation can help identify the most effective treatments for patients with epidermal growth factor receptor (EGFR)-positive lung cancer, ensuring that they receive personalised treatments that target their specific mutation.

In conclusion, mutations in the epidermal growth factor receptor-H773_V774 gene are a major cause of certain types of lung cancer and an important therapeutic target for the treatment of EGFR-positive lung cancer. Targeting this mutation with tyrosine kinase inhibitors improves patient prognosis and personalised medical approaches, providing a valuable strategy in the fight against lung cancer.

Customer Q&As

What is the EGFR-H773_V774insNPH gene and its role in cancer?

A: The EGFR-H773_V774insNPH represents a novel insertion mutation identified in the epidermal growth factor receptor (EGFR) gene. This mutation, located in exon 20, involves the insertion of the sequence "NPH" (asparagine-proline-histidine) between amino acids 773 and 774 of the EGFR protein. This insertion mutation can lead to aberrant EGFR signaling and may contribute to the development of resistance to EGFR-targeted therapies in certain cancers, particularly non-small cell lung cancer (NSCLC).

How does the EGFR-H773_V774insNPH mutation affect EGFR function?

A: The EGFR-H773_V774insNPH mutation affects EGFR function by altering the structure and activity of the kinase domain. This insertion can lead to the formation of a novel activation loop, which enhances the kinase activity of the EGFR protein. As a result, the mutated EGFR can drive uncontrolled cell growth and survival, contributing to the development of cancer.

Are there any therapeutic implications of targeting the EGFR-H773_V774insNPH mutation?

A: Targeting the EGFR-H773_V774insNPH mutation has therapeutic implications for the treatment of cancers that harbor this specific mutation. Inhibitors that can specifically target this mutation, such as osimertinib (Tagrisso), may be effective in treating patients with NSCLC and acquired resistance to first-line EGFR inhibitors. These drugs can lead to tumor regression and improve patient outcomes.

How is the EGFR-H773_V774insNPH mutation detected and diagnosed?

A: The EGFR-H773_V774insNPH mutation is detected through molecular diagnostic techniques such as next-generation sequencing (NGS), which can identify the specific insertion mutation in the EGFR gene. This allows for accurate diagnosis and prognosis of patients with NSCLC and acquired resistance to EGFR inhibitors. Reverse transcription polymerase chain reaction (RT-PCR) and other molecular diagnostic methods can also be used to detect this mutation.

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Customer Reviews

Product development

The Human EGFR-H773_V774insNPH Stable Cell Line-Ba/F3 is a result of meticulous product development, providing researchers with a valuable tool to study the effects of specific EGFR mutations on cell behavior and response to potential therapeutics.

United States

2020-04-06

Knockout efficiency

This cell line demonstrates high knockout efficiency, ensuring that the desired mutation is effectively incorporated into the EGFR gene. This efficiency allows researchers to confidently study the biological consequences of this specific mutation.

Canada

2023-03-19

Genetic stability

The Human EGFR-H773_V774insNPH Stable Cell Line-Ba/F3 exhibits high genetic stability, maintaining the mutation over multiple cell divisions. This stability is crucial for conducting long-term experiments and ensures consistent and reliable results.

Germany

2022-12-05

Cell growth and morphology

The cell line shows robust growth and maintains a normal morphology, which is essential for accurate interpretation of experimental results. The consistent cell growth allows researchers to perform experiments with high reproducibility and reliability.

Canada

2021-07-07

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Human EGFR-H773_V774insNPH Stable Cell Line-Ba/F3

Cell Line Information

Cell Culture Information

Safety and Packaging

Gene Information

Background

Case Study

Applications

Q & A

Customer Reviews

What is the EGFR-H773_V774insNPH gene and its role in cancer?

How does the EGFR-H773_V774insNPH mutation affect EGFR function?

Are there any therapeutic implications of targeting the EGFR-H773_V774insNPH mutation?

How is the EGFR-H773_V774insNPH mutation detected and diagnosed?

Product development

Knockout efficiency

Genetic stability

Cell growth and morphology

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