The GOLM1 Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma line, engineered for targeted disruption of the GOLM1 gene. This polyclonal format provides a heterogeneous pool of knockout cells, enabling researchers to study loss-of-function effects without relying on single-cell clonal expansion. The product serves as a robust tool for interrogating GOLM1-dependent mechanisms in cancer biology, particularly those governing signal transduction, cell adhesion, and metastatic progression.
HT29 is an extensively characterized epithelial cell line established from a primary colorectal adenocarcinoma of a 44-year-old female. It is widely employed as a model system for colorectal cancer and intestinal biology, retaining key oncogenic mutations and epithelial differentiation characteristics. The cell line exhibits moderate tumorigenicity and has been instrumental in dissecting pathways such as EGFR and Wnt signaling, making it a physiologically relevant host for investigating GOLM1 function in colon adenocarcinoma.
GOLM1 encodes a type II Golgi membrane protein that participates in vesicular trafficking, protein glycosylation, and the regulation of cell surface receptor dynamics. Mechanistically, GOLM1 binds directly to EGFR and facilitates its recycling back to the plasma membrane, thereby sustaining ligand-induced activation of EGFR/STAT3 signaling. This engagement prolongs downstream oncogenic cascades, including PI3K/AKT/mTOR and Wnt/??-catenin pathways. Key downstream effectors modulated by GOLM1 include phosphorylated STAT3, AKT, MMP9, N-cadherin, and vimentin, while upstream regulators such as EGF, IL-6, and STAT3 itself can further amplify GOLM1 expression. Interacting partners like Rab1a and Golgi matrix proteins cooperate in mediating these pro-tumorigenic functions.
Within the HT29 cellular context, GOLM1 contributes to phenotypes central to colorectal carcinogenesis, including enhanced proliferation, migration, and invasiveness. Genetic disruption of GOLM1 in these cells offers a clean system to dissect the reliance of EGFR/STAT3-driven malignancy on Golgi-mediated receptor trafficking. This knockout model is particularly pertinent for exploring the interplay between Golgi biology and oncogenic signaling, and for evaluating GOLM1 as a potential therapeutic vulnerability in colorectal and other cancers where it is overexpressed, such as hepatocellular carcinoma and lung cancer.
Researchers can employ the GOLM1 Knockout HT29 Polyclonal Cells in a broad array of functional assays. Commonly used techniques include western blotting to assess changes in total and phosphorylated EGFR, STAT3, and AKT; RT-qPCR to quantify transcript levels of downstream targets like MMP9 and vimentin; and Transwell migration/invasion assays to measure metastatic capacity. Proliferation can be monitored via MTT or CCK-8 assays, while co-immunoprecipitation experiments verify the loss of EGFR-GOLM1 interaction. Immunofluorescence staining enables visualization of Golgi architecture, and xenograft tumor models allow in vivo assessment of tumor growth and dissemination. For additional technical information, please contact Ascent Research.
