The CD59 Knockout HGC-27 Polyclonal Cells product comprises a polyclonal population of HGC-27 human gastric adenocarcinoma cells engineered by CRISPR/Cas9-mediated disruption of the CD59 gene. This knockout model provides a heterogeneous mixture of edited cells, each carrying distinct loss-of-function mutations in CD59, enabling pooled studies of complement regulation and immune evasion in a cell-autonomous gastric cancer background. The polyclonal nature eliminates clonal bias and allows for the assessment of diverse genetic alterations within the same experimental context.
The parental HGC-27 cell line was originally established from the lymph node metastasis of a patient with gastric adenocarcinoma, making it a widely utilized model for studying metastatic gastric cancer. These cells retain key features of gastric adenocarcinoma and are frequently employed in investigations of tumor cell invasion, complement resistance, and immunomodulatory mechanisms relevant to advanced disease.
CD59 encodes a glycosylphosphatidylinositol (GPI)-anchored glycoprotein that serves as a critical inhibitor of the complement membrane attack complex (MAC). Mechanistically, CD59 binds to complement components C8 and C9, preventing C9 polymerization and subsequent MAC assembly, thereby shielding cells from complement-dependent cytotoxicity. Beyond complement regulation, CD59 functions as a T cell co-stimulatory molecule via interaction with CD2, contributing to T cell receptor signaling. Its expression is modulated by inflammatory stimuli, including TNF-alpha and IL-1 beta, through NF-kB and STAT3 transcriptional pathways. In the context of cancer, CD59 cooperates with other complement regulators such as CD55 and CD46 to facilitate immune evasion, and its localization in lipid rafts positions it within signaling platforms that influence tumor cell survival and immune synapse formation.
In HGC-27 gastric adenocarcinoma cells, CD59-mediated complement resistance is particularly relevant to the metastatic phenotype, as tumor cells in the lymph node microenvironment encounter high levels of complement activity. Disruption of CD59 renders these cells susceptible to complement-dependent lysis, making the knockout model valuable for dissecting complement evasion mechanisms intrinsic to metastatic dissemination. Moreover, since CD59 also modulates T cell co-stimulation, its loss may alter tumor-immune cell interactions, providing a tool to study the dual role of complement regulators in tuning both innate and adaptive immune responses in gastric cancer.
This polyclonal CD59 knockout model is ideal for functional studies employing complement-dependent cytotoxicity assays, where loss of protection enhances sensitivity to complement attack, facilitating evaluation of therapeutic antibodies that rely on complement activation. It enables flow cytometry-based phenotypic analysis of MAC deposition and cell death, as well as western blotting and immunofluorescence to verify CD59 ablation. T cell co-stimulation assays can be used to assess the contribution of CD59 to lymphocyte activation. RNA-seq experiments comparing knockout and wild-type populations can uncover transcriptional changes linked to complement resistance and immune evasion. For additional information or customized solutions, please contact Ascent Research.
