This study investigates the effects of cannabidiol (CBD) in combination with photodynamic therapy (PDT) on MCF-7 breast cancer cells. CBD, a non-psychoactive compound from Cannabis sativa, has shown potential in cancer treatment due to its anticancer properties. When combined with PDT, which uses photosensitizers and light to destroy cancer cells, CBD enhances the therapeutic effect. The study used CBD at varying concentrations (1.25–20 μg/mL) and combined it with PDT using hypericin-gold nanoparticle conjugates. Morphological and biochemical assays, including lactate dehydrogenase (LDH) for membrane integrity, adenosine triphosphate (ATP) for cell viability, and trypan blue exclusion for viability, were used to assess the effects of CBD and PDT. The results showed that CBD at higher concentrations (5–20 μg/mL) caused significant cell death, characterized by vacuolization, blebbing, and floating. Biochemical assays indicated increased LDH release, decreased ATP levels, and reduced cell viability, suggesting CBD-induced cell death. Immunofluorescence results indicated that cell death occurred via apoptosis, with markers such as Bax, Bcl-2, cytochrome c, and PARP being expressed. The study concludes that CBD combined with PDT is effective in inducing apoptosis in MCF-7 breast cancer cells. This combination therapy shows promise as a potential treatment for breast cancer due to its ability to enhance the effects of PDT and reduce the need for conventional chemotherapy, which can have severe side effects. The findings suggest that CBD and PDT could be a viable alternative for cancer treatment, particularly in hormone-responsive breast cancer cells. Further research is needed to explore the mechanisms of action and to validate these findings in vivo.This study investigates the effects of cannabidiol (CBD) in combination with photodynamic therapy (PDT) on MCF-7 breast cancer cells. CBD, a non-psychoactive compound from Cannabis sativa, has shown potential in cancer treatment due to its anticancer properties. When combined with PDT, which uses photosensitizers and light to destroy cancer cells, CBD enhances the therapeutic effect. The study used CBD at varying concentrations (1.25–20 μg/mL) and combined it with PDT using hypericin-gold nanoparticle conjugates. Morphological and biochemical assays, including lactate dehydrogenase (LDH) for membrane integrity, adenosine triphosphate (ATP) for cell viability, and trypan blue exclusion for viability, were used to assess the effects of CBD and PDT. The results showed that CBD at higher concentrations (5–20 μg/mL) caused significant cell death, characterized by vacuolization, blebbing, and floating. Biochemical assays indicated increased LDH release, decreased ATP levels, and reduced cell viability, suggesting CBD-induced cell death. Immunofluorescence results indicated that cell death occurred via apoptosis, with markers such as Bax, Bcl-2, cytochrome c, and PARP being expressed. The study concludes that CBD combined with PDT is effective in inducing apoptosis in MCF-7 breast cancer cells. This combination therapy shows promise as a potential treatment for breast cancer due to its ability to enhance the effects of PDT and reduce the need for conventional chemotherapy, which can have severe side effects. The findings suggest that CBD and PDT could be a viable alternative for cancer treatment, particularly in hormone-responsive breast cancer cells. Further research is needed to explore the mechanisms of action and to validate these findings in vivo.