• Origin: The NCI-H1975 cell line was isolated in July 1988 from the lung of a non-smoking female patient with non-small cell lung cancer. These cells can be used in immuno-oncology and lung cancer research.
• Background Information: This gene encodes a protein that facilitates epidermal growth factor (EGF) binding and functions as its receptor. It is integral to the ERBB2-EGFR signaling pathway, mediating cellular responses to amino acids and promoting fibroblast proliferation. Its biological roles include contributing to eyelid development, protein modification, and regulating its own signaling pathway. The protein is localized to various cellular compartments, including the basolateral plasma membrane, endocytic vesicles, and the perinuclear region, with activity primarily on the plasma membrane. It is widely expressed in systems such as the alimentary tract, brain, skin, limbs, and sensory organs. Research links this gene to Coronavirus infectious disease and aortic valve disease, while its human ortholog is associated with numerous pathologies, including colorectal, lung, pancreatic, and prostate cancers, as well as pulmonary tuberculosis. This gene is orthologous to human EGFR.
• Gene targeting strategy: The exogenous promoter CMV and EGFR coding sequence containing L858R, T790M, C797S  mutations were randomly inserted into the B-hEGFR*L858R*T790M*C797S NCI-H1975 cell line.
• Tumorigenicity: Confirmed in B-NDG mice.
• Application: The B-hEGFR*L858R*T790M*C797S NCI-H1975 tumor model can be used for preclinical evaluation of monoclonal antibody drugs and bispecific antibody drugs.
• Notes:

Inoculated cell lines can be suspended with RPMI-1640 stock solution.

Before implementing the project, it is recommended to perform tumor growth experiments. The recommended cell inoculation amount is between 5E5~1E6.

In the experiment, it is necessary to ensure that the number of animals inoculated subcutaneously is at least 1.6 times the actual grouping number.

• Human EGFR was detected on the surface of both NCI-H1975 and B-hEGFR*L858R*T790M*C797S NCI-H1975.

EGFR expression analysis in NCI-H1975 and B-hEGFR*L858R*T790M*C797S NCI-H1975 by flow cytometry. Single cell suspensions from wild-type NCI-H1975 and B-hEGFR*L858R*T790M*C797S NCI-H1975 #2-F02 cultures were stained with anti-human EGFR antibody (Biolegend, 352904).

Subcutaneous tumor growth of B-hEGFR*L858R*T790M*C797S NCI-H1975 cells. B-hEGFR*L858R*T790M*C797S NCI-H1975 (1×10⁶) and wild-type NCI-H1975 cells (1×10⁶) were subcutaneously implanted into B-NDG mice (female, 7-week-old, n=6). Tumor volume and body weight were measured twice a week. (A) Average tumor volume. (B) Body weight. Volume was expressed in mm³ using the formula: V=0.5 × long diameter × short diameter². Results indicate that B-hEGFR*L858R*T790M*C797S NCI-H1975 cells were able to establish tumors in vivo and can be used for efficacy studies. Values are expressed as mean ± SEM.

• Human EGFR was detected on the surface of B-hEGFR*L858R*T790M*C797S NCI-H1975 tumors cells and wild-type NCI-H1975 cells.

EGFR expression was evaluated​​ on B-hEGFR*L858R*T790M*C797S NCI-H1975​​ by flow cytometry. ​​These cells​​ were subcutaneously transplanted into B-NDG mice (n=6). At the end of the experiment, tumor cells were harvested and ​​analyzed for human EGFR expression​​ by flow cytometry. ​​