The HS3ST1 Knockout MCF-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from the MCF-7 human breast adenocarcinoma cell line, carrying a targeted disruption of the HS3ST1 gene. This product provides a loss-of-function model for studying heparan sulfate 3-O-sulfation within hormone-responsive breast cancer cells. The polyclonal format offers a heterogeneous gene-knockout background, avoiding clonal selection artifacts while preserving the epithelial and metastatic characteristics of the parental line.
MCF-7 cells are an ER-positive, PR-positive, HER2-negative epithelial cell line originating from a pleural effusion metastasis. Widely employed to investigate hormone-responsive breast cancer, MCF-7 exhibits estrogen-dependent proliferation and is a key model for tumor growth, survival, and endocrine resistance. The HS3ST1 knockout in this background enables dissection of heparan sulfate sulfation contributions to cancer cell proliferation, migration, and invasion within a well-defined signaling and hormonal context.
HS3ST1 catalyzes the 3-O-sulfation of glucosamine residues in heparan sulfate, generating binding sites for antithrombin III (SERPINC1) and growth factors such as FGF2, HGF, and VEGFA. This modification occurs on heparan sulfate proteoglycans like SDC1 and depends on the sulfate donor PAPS. Transcription of HS3ST1 is regulated by ESR1, FGF2, and TGFB1. Downstream, HS3ST1 activity facilitates FGF2?CFGFR1 signaling and subsequent ERK phosphorylation, linking sulfation patterns to proliferation and migration. These functional relationships position HS3ST1 at a nexus of coagulation, growth factor signaling, and viral entry pathways.
Disruption of HS3ST1 in MCF-7 cells is expected to abolish 3-O-sulfated heparan sulfate, thereby reducing antithrombin III binding and attenuating growth factor signaling through HSPGs. Specifically, impaired FGF2 presentation by SDC1 likely diminishes FGFR1-mediated ERK activation, leading to decreased proliferation and migration. Additionally, reduced signaling by HGF and VEGFA may further suppress invasive behavior. The knockout model thus provides a tool to examine how heparan sulfate fine structure governs breast cancer cell aggressiveness, particularly under estrogen-dependent regulation of HS3ST1 by ESR1.
This polyclonal knockout is suitable for applications including cancer proliferation, metastasis, and heparan sulfate biology. Researchers can validate knockout via Western blot and RT-qPCR, analyze sulfation changes by immunofluorescence and HPLC-MS, and assess functional impact using antithrombin binding, FGF2-dependent proliferation, and transwell migration/invasion assays. Global expression analysis by RNA-seq and phospho-ERK detection after FGF2 stimulation further characterize signaling alterations. The cells also support viral entry studies. For inquiries, please contact Ascent Research.
