Figure 7.

Evaluation of 5FU-induced cytotoxicity, CD44H cells and autophagy in patient tumor-derived 3D organoids. OPSCC patient tumor-derived 3D organoids were treated with indicated concentrations of 5FU for 72 hours. WST1 assays were performed to determine 5FU IC₅₀. (A) Representative dose response curves for primary (P0) and secondary (P1) 3D organoids from patient 23 (left) are shown along with the IC₅₀ and R square values in the table for 3 independent patients as biological replicates (right). Cell morphology was assessed (B) under phase-contrast microscopy and (C) by H&E staining. Scale bar = 100 μm. (D, E) Tumor-derived 3D organoids from 3 independent patients were grown for 7 days followed by treatment with indicated concentrations of 5FU for 72 hours. (D) CD44H cell content and (E) AV content (Cyto-ID) were evaluated by flow cytometry. The representative histograms of patient 23 (left) are shown with the corresponding quantification (right); data of patients 23, 24, and 26 (3 biological replicates) are presented as mean ± SD. (D, E) One-way analysis of variance with multiple comparisons (Tukey) was used to examine statistical significance. *P < .05 vs 0 μM (n = 3) in each patient. not significant (ns) vs 0 μM (n = 3) in each patient. #No technical replicate was available due to the small number of viable cells that were recovered from 3D organoids in patient 24.