The GOLPH3 Knockout HT29 Polyclonal Cells are a heterogeneous population of HT29 human colorectal adenocarcinoma cells that have undergone CRISPR/Cas9-mediated disruption of the GOLPH3 locus. This polyclonal knockout product provides a loss-of-function model to study GOLPH3 oncogenic roles without clonal selection artifacts, suitable for pooled functional studies.
The HT29 cell line, derived from a primary colon adenocarcinoma, carries a TP53 mutation and retains the ability to undergo enterocytic differentiation. These characteristics render HT29 a versatile model for exploring colorectal cancer signaling and intestinal epithelial biology.
GOLPH3 is a Golgi-resident phosphoprotein that links vesicular trafficking to oncogenic pathways. Transcriptionally activated by MYC and stimulated by PI3K/EGF signaling, GOLPH3 interacts with MYO18A and F-actin to maintain Golgi morphology, and binds RPTOR to drive mTORC1 activity. This promotes phosphorylation of S6K and AKT, while also enhancing Wnt/??-catenin transcriptional output. Thus, GOLPH3 couples Golgi dynamics to the MYC??GOLPH3??mTORC1(RPTOR/MTOR)??AKT/S6K cascade and ??-catenin signaling.
In HT29 cells with mutant p53, GOLPH3 knockout impairs Golgi integrity and attenuates mTOR/AKT and Wnt signaling, resulting in reduced proliferation, increased apoptosis, and decreased invasion. This polyclonal model enables precise dissection of Golgi-dependent oncogenic signaling in colorectal cancer, providing a system to probe pathway dependencies and therapeutic targets.
Researchers can assess mTOR/AKT activity via phospho-S6K and phospho-AKT western blotting, and Wnt output by RT-qPCR of ??-catenin target genes. Proliferation, apoptosis, and invasion are measured by MTT, flow cytometry, and transwell assays, while immunofluorescence reveals Golgi disruption. These cells are ideal for drug screening against mTOR/Wnt pathways and Golgi trafficking studies. For technical details, contact Ascent Research.
