title: Cell death mechanistic study of photodynamic therapy against breast cancer cells utilizing liposomal delivery of 5,10,15,20-tetrakis(benzo[b]thiophene) porphyrin.
abstract: 5,10,15,20-Tetrakis(benzo[b]thiophene) porphyrin (BTP) is a newly synthesized hydrophobic photosensitizer with fluorescence quantum yield in toluene: ΦF=0.062. Previously, its limitations in solubility had hindered scientific experimentation regarding its photodynamic effects on cancer cells. By utilizing various compositions of liposomes in order to alter the solubility of BTP, the photocytotoxicity, reactive oxygen species generation, and subcellular localization of the liposomal BTP were identified in this work. DNA fragmentation and high content screening assays were performed in order to shed light on the tumoricidal mechanism of the liposomal photosensitizer. The MTT assay results showed promising results in the irradiation specific PDT activity against MCF-7 cells in all liposomal compositions. Production of ROS was confirmed in the liposomal BTP treated MCF-7 cells after irradiation in a concentration dependent manner. The subcellular localization assays revealed that the localization of BTP was dependent on both the photosensitizer's chemical properties and the properties of the delivery agent encapsulating aforesaid substance. Significant DNA fragmentation was observed in both nucleus localizing liposomal BTP, BTP encapsulated DOPC and DOPE (DOPC-BTP and DOPE-BTP), treated MCF-7 cells. All liposomal-BTPs were successful in inducing mitochondrial permeability transition, an increase in the permeability of the mitochondrial membrane, and activating caspase-3/7. ER localizing BTP were able to significantly increase the cytosolic calcium levels by photodynamic therapy, confirming the photodynamic ability of ER localized BTP to damage the ER membrane. The application of liposomes in delivering a novel hydrophobic photosensitizer, BTP, and photodynamic therapy treatment against MCF-7 cells were successful. It was confirmed that the MCF-7 cell death pathway via photodynamic therapy was altered in a controlled manner by controlling the intracellular localization of the photosensitizer through lipid composition adjustment.
authors.affiliation: College of Pharmacy, Seoul National University, Seoul 151-742, South Korea.
authors.forename: Geewoo
authors.initials: G
authors.lastname: Nam
authors.affiliation: College of Pharmacy, Seoul National University, Seoul 151-742, South Korea.
authors.forename: Sabarinathan
authors.initials: S
authors.lastname: Rangasamy
authors.affiliation: College of Pharmacy, Seoul National University, Seoul 151-742, South Korea.
authors.forename: Hee
authors.initials: H
authors.lastname: Ju
authors.affiliation: College of Pharmacy, Seoul National University, Seoul 151-742, South Korea.
authors.forename: Annie Agnes Suganya
authors.initials: AA
authors.lastname: Samson
authors.affiliation: College of Pharmacy, Seoul National University, Seoul 151-742, South Korea. Electronic address: jmsong@snu.ac.kr.
authors.forename: Joon Myong
authors.initials: JM
authors.lastname: Song
mesh_terms: D001943:Breast Neoplasms / Q000188:drug therapy* / Q000473:pathology; D045744:Cell Line, Tumor; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D046529:Microscopy, Electron, Transmission; D010778:Photochemotherapy; D011838:Radiation-Sensitizing Agents / Q000627:therapeutic use*; D017382:Reactive Oxygen Species / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013347:Subcellular Fractions / Q000378:metabolism
chemical_list: D011838:Radiation-Sensitizing Agents; D017382:Reactive Oxygen Species
keywords: Cell death pathway; High content screening; Liposome; MCF-7; Photodynamic therapy; Subcellular localization