In vitro insights: Investigating Pro-apoptotic potential of capsaicin against HL-60, Human Leukemia cell line

Objective: To investigate the pro-apoptotic potential of capsaicin on Human Leukemia HL-60 cell line as a potent natural chemotherapeutic agent. Methodology: This study was conducted at Forman Christian College, Lahore from January to April-2024. It included culturing HL-60 cell line and its treatment with serial concentrations of capsaicin for calculation of subsequent IC50 values via MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay, expression analysis of gene, comparative analysis of relative gene fold (Intrinsic Biomarkers Caspase-3, Caspase-9) as apoptosis mediator markers followed by RT-qPCR. Cellular apoptotic morphology was confirmed via Hoechst 333258 staining. Results: For HL-60 cell line, the IC50 of capsaicin (16.7uM) showed high gene expression of proapoptotic biomarkers (Caspase-3 and Caspase-9). Hoechst 333258 staining showed prominent apoptotic bodies leading to nuclear fragmentations. Conclusion: Capsaicin had robust pro-apoptotic potential leading to diagnostic efficacy against leukemia HL-60 cell line. Further studies would help in identifying key mechanisms by which capsaicin exhibits anti-apoptotic potential against HL-60 cell lines.