The HS3ST1 Knockout HGC-27 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population of the HGC-27 human gastric carcinoma cell line. This heterogeneous cell pool harbors targeted disruptions of the HS3ST1 gene, encoding heparan sulfate 3-O-sulfotransferase 1, without single-cell cloning. The polyclonal format retains the parental line??s genetic diversity, providing a loss-of-function model that reflects the biological variability of metastatic cancer cells. The CRISPR/Cas9-mediated gene disruption eliminates HS3ST1 function, enabling studies of 3-O-sulfated heparan sulfate-dependent processes.
HGC-27 is a metastatic gastric adenocarcinoma cell line derived from a lymph node metastasis. It displays aggressive proliferation, invasiveness, and molecular alterations typical of advanced gastric cancer, including dysregulated growth factor signaling and altered heparan sulfate proteoglycan expression. This background makes HGC-27 especially relevant for investigating how specific heparan sulfate modifications influence tumor progression, metastasis, and viral susceptibility.
HS3ST1 catalyzes the 3-O-sulfation of glucosamine residues in heparan sulfate, generating rare motifs that serve as high-affinity binding sites for antithrombin III and HSV-1 glycoprotein gD. These modifications potentiate antithrombin??s anticoagulant activity and enable viral entry. Additionally, 3-O-sulfated heparan sulfate promotes FGF2/FGFR1 complex formation and ERK phosphorylation, and it facilitates Wnt3a/LRP6-mediated ??-catenin signaling. Thus, HS3ST1 acts upstream of antithrombin III, FGF2, Wnt3a, and HSV-1 gD, integrating coagulation, growth factor, and pathogen-entry pathways.
In HGC-27 cells, HS3ST1 knockout eliminates 3-O-sulfation, abrogates antithrombin III binding, and impairs the cellular contribution to anticoagulation. The loss of HS3ST1 also attenuates FGF2-induced phospho-ERK signaling and Wnt3a-driven ??-catenin activation, both of which are linked to gastric cancer proliferation and migration. Moreover, the cells lack the primary receptor for HSV-1 gD, rendering them resistant to infection. The polyclonal nature ensures that the knockout population mirrors the heterogeneity of clinical tumors, enhancing the translational value for studying metastasis, cancer signaling, and pathogen interactions.
These cells are suited for a range of assays, including Western blotting and RT-qPCR to confirm HS3ST1 disruption, heparan sulfate disaccharide analysis to assess sulfation changes, and functional tests such as antithrombin activation, HSV-1 entry, FGF2-stimulated phospho-ERK Western blotting, Wnt luciferase reporter, and migration assays. Applications span cancer biology, viral infection mechanisms, anticoagulation research, and heparan sulfate modification studies. For technical inquiries, please contact Ascent Research.
