Panoply™ Human EGFR Over-expressing Stable Cell Line

Name: Panoply™ Human EGFR Over-expressing Stable Cell Line
SKU: CSC-SC004772
Availability: InStock

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

Cat. No. : CSC-SC004772

Host Cell : HEK293 (CHO and other cell types are also available) Size : >1x10⁶ frozen cells/vial

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Cell Line Information

Cell Culture Information

Safety and Packaging

Gene Information

Cat. No.	CSC-SC004772
Description	Using Creative Biogene's proprietary lentiviral vectors, we subclone the target gene into lentivector, generate the lentivirus particles, sequentially infect the cell line HEK293 (other cell types are also available according to your requirements), and select the clones constantly expressing target gene at high level.
Target Gene	EGFR
Gene Species	Homo sapiens (Human)
Host Cell	HEK293 (CHO and other cell types are also available)
Host Cell Species	Species varies
Applications	1. Gene expression studies 2. Signaling pathway research 3. Drug screening and toxicology 4. Disease research
Size	2 × 10^6 cells / vial
Stability	Validated for at least 10 passages
Quality Control	Negative for bacteria, yeast, fungi and mycoplasma.
Storage	Liquid nitrogen
Shipping	Dry Ice

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

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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Cetuximab resistance is a major obstacle to the treatment of epidermal growth factor receptor (EGFR) overexpressing cancers, including triple-negative breast cancer (TNBC). MicroRNA (miRNA)-155-5p is highly expressed in TNBC cells. Therefore, researchers investigated whether downregulating miR-155-5p could enhance the antitumor effect of cetuximab in TNBC cells. First, they constructed EGFR-overexpressing cell lines (MDA-MB-231 and MDA-MB-468). The inhibitory effect of cetuximab on the proliferation and migration of EGFR-overexpressing MDA-MB-468 cells was enhanced after transfection with a miR-155-5p antagonist, while miR-155-5p knockdown enhanced the pro-apoptotic effect of cetuximab on EGFR-overexpressing MDA-MB-468 cells. Furthermore, luciferase reporter gene assays showed that gasdermin E (GSDME) is a direct binding target of miR-155-5p. Cetuximab in combination with a miR-155-5p antagonist promotes pyroptosis in EGFR-overexpressing MDA-MB-468 cells by upregulating GSDME-N and cleaved caspase-1. In vivo experiments confirmed that downregulation of miR-155-5p enhanced the antitumor effect of cetuximab in MDA-MB-468 xenograft models and EGFR-overexpressing TNBC cells, through the induction of apoptosis and pyroptosis. Therefore, cetuximab combined with a miR-155-5p antagonist may be a novel therapeutic strategy for TNBC.

Here, researchers used flow cytometry to detect the cytotoxic effects of cetuximab combined with a miR-155-5p antagonist on EGFR-overexpressing MDA-MB-468 cells. The results showed that cell death occurred in apoptosis (quadrants Q2 and Q3), necrosis, or pyroptosis (quadrants Q1 and Q2). As shown in Figures 1A and 1B, 10 nM cetuximab significantly induced apoptosis in EGFR-overexpressing MDA-MB-468 cells, while downregulation of miR-155-5p significantly enhanced the induction of necrosis or pyroptosis in EGFR-overexpressing TNBC cells. Furthermore, compared to the cetuximab monotherapy group, the expression of Bax and cleaved caspase-3 was increased in EGFR-overexpressing MDA-MB-468 cells treated with cetuximab combined with a miR-155-5p antagonist. However, compared with the cetuximab monotherapy group, the cetuximab plus miR-155-5p treatment group showed decreased Bcl-2 levels in EGFR-overexpressing MDA-MB-468 cells (Figures 1C-F). These data suggest that downregulation of miR-155-5p enhances the cytotoxic effect of cetuximab on EGFR-overexpressing MDA-MB-468 cells.

Figure 1. Downregulation of miR-155-5p enhances the pro-apoptotic effect of cetuximab in TNBC cells. (Xu W, et al., 2021)

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