The ANLN Knockout KYSE-30 Polyclonal Cells are a heterogeneous population of human esophageal squamous cell carcinoma cells (KYSE-30) with CRISPR/Cas9-mediated disruption of the anillin (ANLN) gene. This polyclonal pool contains a mixture of edited alleles, providing a robust loss-of-function model without clonal selection. The diverse indel mutations abrogate ANLN protein expression, enabling assessment of collective ANLN deficiency effects on cellular phenotypes. Designed for direct in vitro use, these cells offer a powerful tool for investigating ANLN-dependent mechanisms in cancer biology.
The KYSE-30 host cell line originates from a well-differentiated invasive esophageal squamous cell carcinoma (ESCC) of the human esophagus. Widely employed to study ESCC pathogenesis, this cell line retains esophageal epithelial features such as cytokeratin expression and exhibits robust proliferation and motility. It serves as a clinically relevant model for exploring cytoskeletal regulation and cell division pathways dysregulated in esophageal cancer.
Anillin (ANLN) is a multidomain scaffold protein that binds actin, myosin II, and septins, and is essential for cytokinesis and cell migration. ANLN is a central hub in RhoA signaling: recruited by active RhoA and Ect2 to the equatorial cortex, it scaffolds the contractile ring and links the plasma membrane to the actomyosin cytoskeleton. Transcription of ANLN is positively regulated by E2F1 and FOXM1, downstream of the PI3K/AKT/mTOR cascade. ANLN organizes actin dynamics via the RhoA-ROCK-LIMK-cofilin pathway, ensuring proper furrow ingression. Its loss disrupts these interactions, causing cytokinetic failure and reduced motility, underscoring its multifaceted role in cancer.
In esophageal cancer, ANLN overexpression correlates with aggressive proliferation, migration, and invasion. This polyclonal ANLN knockout in KYSE-30 cells enables interrogation of anillin’s oncogenic functions. ANLN disruption is predicted to impair contractile ring integrity, generating multinucleate cells and mitotic defects, while attenuating actin-dependent migration. The model elucidates how ANLN integrates PI3K/AKT, E2F1, and FOXM1 signals to drive ESCC malignancy, and is suited for studying the RhoA-anillin axis in cytokinesis failure and aneuploidy.
These knockout cells are applicable to broad functional studies, including Western blot-based confirmation of ANLN loss, immunofluorescence for cytokinetic defects (multinucleation, cleavage furrow aberration), wound healing and transwell assays for migration/invasion, MTT proliferation assays, and flow cytometric cell cycle analysis for G2/M arrest. RNA-seq can reveal transcriptomic changes. The model supports drug target validation and fundamental cell division research. For technical inquiries, contact Ascent Research.
