The CCDC71L Knockout Huh-7 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Huh-7 human hepatocellular carcinoma cell line. This product provides a loss-of-function model for the CCDC71L gene, which encodes a coiled-coil domain protein involved in oncogenic signaling. The polyclonal format ensures a heterogeneous, gene-disrupted population suitable for pooled functional studies, where the knockout of CCDC71L is achieved without clonal selection. This enables robust analysis of gene function while mitigating clonal artifacts.
The Huh-7 host cell line is a well-differentiated hepatocellular carcinoma line with a p53 mutation, widely employed in liver cancer research and hepatitis C virus (HCV) replication studies. Huh-7 cells exhibit robust hepatic metabolic activity, continuous proliferation, and tumorigenic potential, making them a relevant model for hepatocellular carcinoma pathobiology. The p53-deficient background further accentuates the dependence on alternative survival and proliferation signals, such as those mediated by PI3K/AKT, thus providing a sensitive context for interrogating CCDC71L function.
CCDC71L is a coiled-coil domain-containing protein that interacts with PI3K, enhancing AKT phosphorylation downstream of growth factor receptors such as EGFR and MET. Upon stimulation by EGF or HGF, CCDC71L facilitates the recruitment and activation of PI3K, leading to PDK1-mediated phosphorylation of AKT at Thr308. Activated AKT subsequently phosphorylates GSK-3??, stabilizing ??-catenin and promoting TCF/LEF-mediated transcription of targets like Cyclin D1 and Survivin. Additionally, AKT signaling upregulates MMP2 and MMP9 expression, fostering epithelial-mesenchymal transition and invasiveness, while Bcl-2 induction supports survival. The protein also interfaces with the Wnt/??-catenin pathway through ??-catenin interaction, amplifying proliferative and survival cues. In the context of c-Myc transcriptional regulation, CCDC71L integrates mitogenic and survival signals to drive hepatocellular carcinoma progression.
Disruption of CCDC71L in Huh-7 cells abrogates critical oncogenic signals, particularly those dependent on PI3K/AKT hyperactivation. In the p53-mutant background, CCDC71L knockout is expected to reduce AKT phosphorylation, leading to decreased expression of Cyclin D1 and Survivin, thereby impairing cell cycle progression and enhancing apoptotic sensitivity. Moreover, the suppression of MMP2 and MMP9 attenuates migratory and invasive capacities, which are hallmarks of metastatic hepatocellular carcinoma. This polyclonal knockout population provides a physiologically relevant platform to dissect the contribution of CCDC71L to tumor cell proliferation, survival, and metastasis, circumventing the compensatory mechanisms often observed in p53-competent models. Consequently, it serves as an invaluable tool for studying PI3K/AKT-driven hepatocarcinogenesis and for evaluating therapeutic strategies targeting this axis.
Researchers can utilize these cells to validate CCDC71L as a therapeutic target for hepatocellular carcinoma, employing assays such as CCK-8 cell viability analysis, phospho-AKT Western blotting, and Transwell migration/invasion assays. The knockout model is also suitable for flow cytometric apoptosis detection using Annexin V/PI staining and for in vivo tumorigenicity studies via xenograft models. Furthermore, the cells provide a system for screening small-molecule inhibitors targeting the PI3K/AKT pathway or downstream effectors, and for functional genomics screens aimed at uncovering synthetic lethal interactions. The CCDC71L Knockout Huh-7 Polyclonal Cells offer a robust and versatile resource for advancing liver cancer research. For additional information, please contact Ascent Research.
