Hypericin Photodynamic Therapy Induces Cytotoxicity and Modulates Cytokine Secretion in MCF-7 Breast Cancer Cells

<b>Background/Aim</b>: Photodynamic therapy uses a photosensitizer and light to generate reactive oxygen species that kill tumor cells and can shift inflammatory signaling. Hypericin is a potent photosensitizer, but its immunomodulatory impact in breast cancer needs clarification. We evaluated the phototoxic and cytokine-modulating effects of hypericin-mediated photodynamic therapy in MCF-7 human breast adenocarcinoma cells. This study examines how HYP-PDT affects MCF-7 breast cancer cells by assessing viability and cytokine secretion to guide the development of targeted, immune-enhancing PDT protocols. <b>Methods</b>: MCF-7 cells were incubated with hypericin at 0, 0.125, 0.25, 0.5, or 1 μM, then exposed to light doses of 0, 1, 2, or 5 J/cm². Viability was measured 24 h later by MTT; selected conditions were also assessed by Trypan Blue. Cell supernatants collected after sublethal treatment were analyzed for IL-6, IL-8, IL-10, and TNF-α using a multiplex immunoassay. Experiments were repeated four times. Statistical analyses followed the study's plan for group comparisons. <b>Results</b>: At 1 J/cm², MTT values did not differ from matched dark controls across hypericin concentrations. At 2 and 5 J/cm², some conditions showed increased MTT signal relative to controls, indicating higher metabolic activity; Trypan Blue performed at 0 J/cm² showed a concentration-dependent reduction in viability with hypericin. Hypericin-PDT decreased IL-6 and IL-8 concentrations and increased TNF-α in MCF-7 supernatants. No statistically significant changes were detected for IL-10. <b>Conclusions</b>: Hypericin-PDT altered inflammatory readouts in MCF-7 cells, with reductions in IL-6 and IL-8 and an increase in TNF-α, consistent with a pro-inflammatory shift. Viability results suggest condition-dependent changes in metabolic activity or survival effects that warrant confirmation with matched cell counts across all light doses. These findings support further standardized dosimetry and multi-line validation of hypericin-PDT in breast cancer models.