The KDM5B Knockout CAL-27 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population with targeted disruption of KDM5B, the gene encoding the H3K4me2/3 demethylase. This heterogeneous knockout pool enables pooled loss-of-function studies without clonal bias, making it ideal for robust phenotypic and biochemical analyses in oral squamous cell carcinoma research.
Derived from the CAL-27 cell line, this model originates from a TP53-mutant tongue squamous cell carcinoma with aggressive and invasive properties. CAL-27 is a widely used model for studying OSCC progression, metastasis, and therapeutic resistance, providing a clinically relevant epithelial background for investigating epigenetic drivers of malignancy.
KDM5B functions as a transcriptional repressor by removing H3K4me2/3 marks at gene promoters, and it complexes with HDAC1/2, NuRD, Sin3B, and CoREST. Its expression is activated by upstream factors including MYC, HIF1A, STAT3, and TGF-?? signaling, while it is post-transcriptionally repressed by miR-137 and miR-29. KDM5B silences tumor suppressor genes such as p16INK4a (CDKN2A), p21 (CDKN1A), p27, PTEN, and HOXA5, and concomitantly promotes cell cycle progression and stemness by suppressing pro-apoptotic and epithelial markers.
In CAL-27 cells, KDM5B knockout blocks its demethylase activity, resulting in H3K4 hypermethylation, reactivation of silenced tumor suppressors, and impaired proliferation, survival, and migration. The polyclonal pool captures the loss-of-function phenotypes, including reduced cancer stem cell maintenance and reversal of EMT, as evidenced by altered expression of E-cadherin and vimentin. These cells thus provide a powerful system to elucidate KDM5B-dependent oncogenic mechanisms and the interplay between epigenetic regulation and TP53 mutation status in OSCC.
Researchers can apply these cells in transcriptomic (RNA-seq) and epigenomic (ChIP-qPCR) profiling, coupled with Western blot detection of H3K4me3, p21, and p16. Functional assays such as MTT/CCK8 proliferation, colony formation, flow cytometry for cell cycle/apoptosis, and transwell migration enable comprehensive phenotypic assessment. The knockout model is also suitable for drug sensitivity screening to identify epigenetic therapies or overcome resistance. For additional support or custom projects, please contact Ascent Research.
