The ECE1 Knockout HGC-27 Polyclonal Cells constitute a CRISPR/Cas9-mediated polyclonal knockout cell population designed for loss-of-function studies of the ECE1 gene in a human gastric carcinoma context. This product provides a heterogeneous pool of edited HGC-27 cells, enabling robust investigation of ECE1-dependent processes without clonal selection artifacts. The targeted disruption of the ECE1 locus abrogates expression of functional endothelin-converting enzyme-1, establishing a valuable model for interrogating endothelin signaling and its contribution to cancer cell biology.
The HGC-27 cell line is a well-characterized epithelial model derived from a lymph node metastasis of human gastric adenocarcinoma. These cells exhibit a metastatic phenotype and are widely employed in studies of gastric cancer progression, invasion, and tumor microenvironment interactions. Their adherent growth and stable karyotype make them suitable for genetic manipulation and downstream phenotypic analyses, including migration assays and signaling pathway evaluation.
ECE1 encodes a membrane-bound metalloprotease that processes big endothelin-1, -2, and -3 into bioactive endothelin peptides. Endothelin-1 signals through G protein-coupled receptors EDNRA and EDNRB, triggering G??q-dependent PLC/PKC/Ca2? cascades and activating MAPK/ERK and PI3K/Akt pathways. Under hypoxia or inflammation, transcription factors HIF1??, AP-1, and NF-??B upregulate ECE1 expression in response to upstream cues such as TGF-?? and TNF-??. Downstream targets of this signaling include c-FOS, c-JUN, and various cell cycle progression genes. Thus, ECE1 integrates environmental stress signals with mitogenic and vasoregulatory outputs.
In HGC-27 gastric cancer cells, disruption of ECE1 abolishes the conversion of big endothelin-1 to mature endothelin-1, effectively disrupting autocrine and paracrine signaling loops that promote tumorigenic phenotypes. This loss attenuates downstream phosphorylation of ERK and Akt, impairing cell cycle progression, survival, and invasive motility. Consequently, the knockout model reveals how ECE1-dependent endothelin production contributes to gastric cancer aggressiveness and offers a platform to study the impact of endothelin signaling on tumor angiogenesis and the metastatic microenvironment. Given the link between endothelin dysregulation and hypertension, these cells also enable exploration of cancer?Ccardiovascular disease intersections.
Researchers can deploy these polyclonal knockout cells in quantitative Western blot analysis of phospho-ERK and phospho-Akt, ELISA measurements of endothelin-1 secretion, and RT-qPCR profiling of downstream target transcripts. Functional assays including cell proliferation, migration, and Matrigel invasion can be paired with endothelin receptor antagonists or pathway inhibitors to validate therapeutic targets. Co-immunoprecipitation experiments dissecting ECE1 interaction with big endothelin substrates and immunofluorescence localization studies further characterize the molecular apparatus. This versatile cell population advances ECE1 research in oncology, vascular biology, and preclinical drug screening. Contact Ascent Research for technical support and custom applications.
