Journal: Cancer Management and Research
Article Title: Saikosaponin D Induced Apoptosis and Anti-Tumor Effect Enhancing Chemotherapy Sensitivity of Gemcitabine in Patient-Derived Cholangiocarcinoma Organoids
doi: 10.2147/CMAR.S564565
Figure Lengend Snippet: SSd enhancing chemotherapy sensitivity of gemcitabine in patient-derived cholangiocarcinoma organoids via apoptosis induction. ( A ) TUNEL staining was employed to visualize DNA fragmentation (indicated by brown-stained nuclei) in SSd-treated organoids. Scale bar: 200µm. ( B ) Quantitative analysis of TUNEL-positive cells. Data are presented as mean ± SEM (n=3). Statistical significance was determined by two-way ANOVA (factors: treatment and organoid line) with Tukey’s multiple comparisons test. The analysis revealed a significant main effect of treatment (p < 0.001). For clarity, only select key comparisons are denoted: ****p < 0.0001. ( C ) Caspase-3 activation was assessed by GreenNuc™ fluorescence (green) in treatment groups. An intensified signal in the combination group demonstrated synergistic apoptosis induction. Scale bar: 200µm. ( D ) JC-1 assay was utilized to evaluate mitochondrial depolarization, which was manifested as a shift from red to green fluorescence. SSd treatment reduced red fluorescence (intact mitochondria), with further reduction in the combination group. Scale bar: 200μm.
Article Snippet: Caspase-3 activity detection: caspase-3 activity in the organoid samples was detected using the Caspase-3 Fluorescence Detection Kit (Beyotime, C1073S).
Techniques: Derivative Assay, TUNEL Assay, Staining, Activation Assay, Fluorescence