Research Paper Volume 16, Issue 1 pp 627—647

Targeting of FSP1 regulates iron homeostasis in drug-tolerant persister head and neck cancer cells via lipid-metabolism-driven ferroptosis

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Figure 2. Cisplatin DTP HNSCC cancer cell line generation and validation. (A) Cell line expression analysis to optimize the experimental parameters used HSC3; HSC4 was the control. (B) HSC3 parental cells were treated with 5 μg/mL cisplatin for 9 days, removed from cisplatin for 28 days, and then reincubated with cisplatin for 9 days to generate a stable DTP cell line. (C) Relative gene expression assay indicated higher expression of FSP1, GPX4, and ACSL4 in cisplatin-DTP cell lines. (D) HSC3-P was compared with parental counterpart in terms of sensitivity to cisplatin treatment for validation. (E) Immunofluorescence staining analysis of the expression of FSP1 and ACSL4 in the DTP HNSCC cancer cell line. (F) Diameter of tumor spheres and (G) relative gene expression of FSP1, GPX4, and ACSL4 with and without shFSP1 transfection. (H) Downregulation of lipid components following FSP1 suppression. (*p < 0.05, **p < 0.01, ***p < 0.001). Scale bar: 5 μm.