The Comparison of Anticancer Efficiency of Methylene Blue- and Aluminum Phthalocyanine-Mediated Sonophotodynamic Therapy on Prostate Cancer Cells In Vitro

Abstract

Prostate cancer ranks highest in male diagnoses and second in cancer-related deaths. Conventional treatments necessitate exploration of new modalities due to their side effects. Sonophotodynamic therapy (SPDT) represents a potential anticancer approach that integrates both sonodynamic and photodynamic therapies to improve the efficacy of cancer treatments. This study aims to evaluate and compare the mechanisms and anticancer efficacy of photodynamic therapy, sonodynamic therapy, and SPDT using methylene blue (MB) and aluminum phthalocyanine (AlPc) in the androgen-sensitive and androgen-insensitive prostate cancer cell lines. Cells were cultivated using different concentrations of MB and AlPc, followed by the exposure to ultrasound and/or light irradiation. Cell metabolic activity was assessed using the MTT assay, which evaluates mitochondrial enzyme function as an indicator of viability rather than clonogenic survival. Additionally, apoptosis was evaluated using Hoechst staining and Western blot analysis of apoptotic proteins, while reactive oxygen species (ROS) and antioxidant levels were determined through biochemical assays. Results showed significant proliferation inhibition, with SPDT exhibiting the highest efficacy. MB demonstrated superior efficiency compared to AlPc. The treatment groups displayed a greater quantity of apoptotic cells, indicating elevated levels of caspase-3, caspase-8, PARP, and Bax proteins, whereas levels of caspase-9 and Bcl-2 were lower compared to the control groups. Additionally, the treatments resulted in increased levels of ROS and malondialdehyde, while antioxidant activities were diminished. In summary, SPDT mediated by MB and AlPc presents promising potential for treating prostate cancer and may significantly contribute to apoptotic mechanisms.