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- 2019
Hypocrellin A-based photodynamic action induces apoptosis in A549 cells through ROS-mediated mitochondrial signaling pathwayDOI: 10.1016/j.apsb.2018.12.004 Keywords: ACN, acetonitrile, CLSM, confocal laser scanning confocal microscopy, DCFH-DA, 2′,7′-dichlorofuorescin diacetate, DMEM, Dulbecco？s modified Eagle？s medium, Dox, doxorubicin, ECL, enhanced chemiluminescence, FCCP, carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone, FDR, false discovery rate, GO, gene ontology, HA, hypocrellin A, HRP, horseradish peroxidase, IAA, iodoacetamide, IKK, IκB kinase complex, iTRAQ, isobaric tag for relative and absolute quantitation, JC-1, 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethyl-benzimidazolcarbocyanine iodide, MPT, mitochondrial permeability transition, MMP, mitochondrial membrane potential, NAC, N-acetyl-l-cysteine, ROS, reactive oxygen species, OCR, oxygen consumption rate, PDT, photodynamic therapy, PI, propidium iodide, PS, photosensitizer, SCX, strong cation exchange, TCM, traditional Chinese medicinal, TEM, transmission electron microscope, TFA, trifluoroacetic acid, UA, urea, z-LEHD-fmk, z-Leu-Glu(OMe)-His-Asp(OMe)-fluoromethylketone, z-VAD-fmk, z-Val-Ala-Asp-fluoromethylketone, z-IETD-fmk, z-Ile-Glu-Asp-fluoromethylketone Hypocrellin A, Photodynamic therapy, Reactive oxygen species, Proteomic, LC–MS/MS, iTRAQ Abstract: Over recent decades, many studies have reported that hypocrellin A (HA) can eliminate cancer cells with proper irradiation in several cancer cell lines. However, the precise molecular mechanism underlying its anticancer effect has not been fully defined. HA-mediated cytotoxicity and apoptosis in human lung adenocarcinoma A549 cells were evaluated after photodynamic therapy (PDT). A temporal quantitative proteomics approach by isobaric tag for relative and absolute quantitation (iTRAQ) 2D liquid chromatography with tandem mass spectrometric (LC–MS/MS) was introduced to help clarify molecular cytotoxic mechanisms and identify candidate targets of HA-induced apoptotic cell death. Specific caspase inhibitors were used to further elucidate the molecular pathway underlying apoptosis in PDT-treated A549 cells. Finally, down-stream apoptosis-related protein was evaluated. Apoptosis induced by HA was associated with cell shrinkage, externalization of cell membrane phosphatidylserine, DNA fragmentation, and mitochondrial disruption, which were preceded by increased intracellular reactive oxygen species (ROS) generations. Further studies showed that PDT treatment with 0.08 μmol/L HA resulted in mitochondrial disruption, pronounced release of cytochrome c, and activation of caspase-3, -9, and -7. Together, HA may be a possible therapeutic agent directed toward mitochondria and a promising photodynamic anticancer candidate for further evaluation
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