Journal: Frontiers in Pharmacology
Article Title: Genome-wide CRISPR screen identified NEK6 as a determinant of sensitivity to CDK4/6 inhibitor in endometrial cancer
doi: 10.3389/fphar.2025.1725886
Figure Lengend Snippet: NEK6 knockout enhanced the efficiency of CDK4/6 inhibitor-induced DNA damage repair and apoptosis. (A) DNA damage detected by comet assay in HEC-1A cells (siNC or si-NEK6) treated with indicated concentration of palbociclib. Scale bar, 50 μm. Quantification of DNA in the tail from three independent experiments is shown as mean ± s.d. (B) Immunofluorescence staining of γ-H2AX in sh-NC and sh-NEK6 HEC-1A and Ishikawa cells treated with or without palbociclib. Scale bar = 10 μm. (C) Western blot analysis of phosphorylated H2AX and RAD51 in HEC-1A and ishikawa endometrial cancer cells treated as indicated. (D) Knockdown of NEK6 induced apoptosis significantly upon palbociclib treatment in HEC-1A evaluated by flow cytometry analysis. Data were presented as the mean ± s.d. of values obtained in 3 independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant.
Article Snippet: The CDK4/6 inhibitors (abemaciclib, palbociclib, ribociclib) and the specific NEK6 inhibitor ZINC05007751 was purchased from MedChem Express (Shanghai, China).
Techniques: Knock-Out, Single Cell Gel Electrophoresis, Concentration Assay, Immunofluorescence, Staining, Western Blot, Knockdown, Flow Cytometry