The HPSE Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HCT 116 human colorectal carcinoma line, offering constitutive disruption of the heparanase (HPSE) gene. This polyclonal pool contains a heterogeneous mix of loss-of-function alleles, making it ideal for studying HPSE-dependent phenotypes without the confounding effects of clonal selection. The model enables researchers to investigate heparanase biology in a background faithful to epithelial colorectal adenocarcinoma.
HCT 116 is a well-characterized epithelial colorectal adenocarcinoma cell line harboring a KRAS G13D mutation and a stabilizing CTNNB1 (??-catenin) mutation, and displaying microsatellite instability-high (MSI-H) status. These oncogenic lesions lead to constitutive activation of MAPK and Wnt/??-catenin signaling, endowing the cells with rapid proliferation, invasive capacity, and resistance to apoptosis. The line is extensively used in preclinical studies of colon cancer metastasis, tumor microenvironment interactions, and drug response.
The HPSE gene encodes an endo-??-glucuronidase that cleaves heparan sulfate side chains from proteoglycans such as perlecan (HSPG2) and syndecans, reshaping the extracellular matrix. This enzymatic action releases matrix-bound growth factors, including VEGF-A, FGF2, and HGF, which subsequently bind VEGFR and FGFR to trigger PI3K/Akt and MAPK signaling. HPSE transcription is upregulated by EGR1, NF-??B, p53, and HIF-1?? in response to TNF-??, IL-1??, or hypoxia. Beyond growth factor mobilization, heparanase promotes the expression of MMP-9 and COX-2, facilitates exosome secretion enriched with CD63, and induces autophagy through LC3 processing. These activities promote cell invasion, angiogenesis, and survival.
In the HCT 116 background, the coexistence of constitutively active KRAS and ??-catenin with HPSE-driven ECM remodeling creates a highly permissive environment for tumor progression. Heparanase-mediated release of VEGF and FGF2 reinforces angiogenic switching and autocrine growth stimulation, while activation of Akt and MAPK pathways attenuates anoikis and fosters chemoresistance. Disrupting HPSE in this context can dampen invasive behavior, reduce angiogenic factor secretion, and impair exosome-based intercellular communication, providing a powerful system to elucidate molecular drivers of colorectal cancer metastasis and to model therapeutic vulnerabilities.
Applications include western blotting and RT-qPCR to confirm HPSE depletion and assess downstream targets, Boyden chamber invasion and wound healing assays to quantify migration and invasiveness, and VEGF ELISA or endothelial tube formation to evaluate angiogenic output. Autophagy flux can be monitored by LC3 immunoblotting, and exosome isolation allows characterization of secreted vesicles. Signaling perturbations are interrogated by phospho-MAPK and phospho-Akt analysis. Additionally, drug sensitivity testing with 5-FU or oxaliplatin permits exploration of HPSE??s contribution to treatment resistance. For further information, please contact Ascent Research.
