-
Cellular Physiology and Biochemistry :... 2015Photodynamic therapy (PDT) is a promising noninvasive technique, which has been successfully applied to the treatment of human cancers. Studies have shown that the Bcl-2...
BACKGROUND/AIMS
Photodynamic therapy (PDT) is a promising noninvasive technique, which has been successfully applied to the treatment of human cancers. Studies have shown that the Bcl-2 family proteins play important roles in PDT-induced apoptosis. However, whether Bcl-2-interacting mediator of cell death (Bim) is involved in photodynamic treatment remains unknown. In this study, we attempt to determine the effect of Bim on Photofrin photodynamic treatment (PPT)-induced apoptosis in human lung adenocarcinoma ASTC-a-1 cells.
METHODS
The translocation of Bim/Bax of the cells were monitored by laser confocal scanning microscope. The levels of Bim protein and activated caspase-3 in cells were detected by western blot assay. Caspase-3 activities were measured by Caspase-3 Fluorogenic Substrate (Ac-DEVD-AFC) analysis. The induction of apoptosis was detected by Hoechst 33258 and PI staining as well as flow cytometry analysis. The effect of Bim on PPT-induced apoptosis was determined by RNAi.
RESULTS
BimL translocated to mitochondria in response to PPT, similar to the downstream pro-apoptotic protein Bax activation. PPT increased the level of Bim and activated caspase-3 in cells and that knockdown of Bim by RNAi significantly protected against caspase-3 activity. PPT-induced apoptosis were suppressed in cells transfected with shRNA-Bim.
CONCLUSION
We demonstrated the involvement of Bim in PPT-induced apoptosis in human ASTC-a-1 lung adenocarcinoma cells and suggested that enhancing Bim activity might be a potential strategy for treating human cancers.
Topics: Apoptosis; Apoptosis Regulatory Proteins; Bcl-2-Like Protein 11; Blotting, Western; Caspase 3; Cell Line, Tumor; Dihematoporphyrin Ether; Humans; Lasers; Membrane Proteins; Microscopy, Electron, Scanning; Mitochondria; Photochemotherapy; Photosensitizing Agents; Proto-Oncogene Proteins; RNA Interference; RNA, Small Interfering; bcl-2-Associated X Protein
PubMed: 25791936
DOI: 10.1159/000373968 -
Potassium Iodide Potentiates Broad-Spectrum Antimicrobial Photodynamic Inactivation Using Photofrin.ACS Infectious Diseases Apr 2017It is known that noncationic porphyrins such as Photofrin (PF) are effective in mediating antimicrobial photodynamic inactivation (aPDI) of Gram-positive bacteria or...
It is known that noncationic porphyrins such as Photofrin (PF) are effective in mediating antimicrobial photodynamic inactivation (aPDI) of Gram-positive bacteria or fungi. However, the aPDI activity of PF against Gram-negative bacteria is accepted to be extremely low. Here we report that the nontoxic inorganic salt potassium iodide (KI) at a concentration of 100 mM when added to microbial cells (10/mL) + PF (10 μM hematoporphyrin equivalent) + 415 nm light (10 J/cm) can eradicate (>6 log killing) five different Gram-negative species (Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Proteus mirabilis, and Acinetobacter baumannii), whereas no killing was obtained without KI. The mechanism of action appears to be the generation of microbicidal molecular iodine (I/I) as shown by comparable bacterial killing when cells were added to the mixture after completion of illumination and light-dependent generation of iodine as detected by the formation of the starch complex. Gram-positive methicillin-resistant Staphylococcus aureus is much more sensitive to aPDI (200-500 nM PF), and in this case potentiation by KI may be mediated mainly by short-lived iodine reactive species. The fungal yeast Candida albicans displayed intermediate sensitivity to PF-aPDI, and killing was also potentiated by KI. The reaction mechanism occurs via singlet oxygen (O). KI quenched O luminescence (1270 nm) at a rate constant of 9.2 × 10 M s. Oxygen consumption was increased when PF was illuminated in the presence of KI. Hydrogen peroxide but not superoxide was generated from illuminated PF in the presence of KI. Sodium azide completely inhibited the killing of E. coli with PF/blue light + KI.
Topics: Anti-Bacterial Agents; Candida albicans; Dihematoporphyrin Ether; Drug Synergism; Gram-Negative Bacteria; Methicillin-Resistant Staphylococcus aureus; Microbial Sensitivity Tests; Photosensitizing Agents; Potassium Iodide
PubMed: 28207234
DOI: 10.1021/acsinfecdis.7b00004 -
Chang Gung Medical Journal 2008The object of the present study was to evaluate the feasibility of photodynamic therapy (PDT) for malignant melanomas through in vivo and in vitro processes.
BACKGROUND
The object of the present study was to evaluate the feasibility of photodynamic therapy (PDT) for malignant melanomas through in vivo and in vitro processes.
METHODS
Photofrin (porfimer sodium) was evaluated through in vitro processes with human malignant melanoma cells (MMCs). The in vitro absorption and photosensitizing activity of Photofrin was examined in an MMC culture system. The in vivo biological activity of Photofrin applied to subcutaneous implanted melanoma (SIM) in nude mice and exposed to different total light dosages at 630 nm was studied by determining the destruction of the tumors. Subcelluar localization and binding were observed under a fluorescent confocal microscope.
RESULTS
MMCs incubated with Photofrin at a concentration of about 3.5 microg/ml and exposed to laser light at 630 nm with a power density of 100 mW/cm2, showed 50% cell killing. An electron microscopic study demonstrated significant destruction of the target after PDT.
CONCLUSION
Detection of the photosensitizer Photofrin was localized and its distribution fully observed. PDT-Photofrin has the capability to destroy MMCs through in vitro and in vivo SIM treatment.
Topics: Animals; Antineoplastic Combined Chemotherapy Protocols; Dihematoporphyrin Ether; Feasibility Studies; Melanoma; Mice; Mice, Nude; Photochemotherapy; Photosensitizing Agents; Tumor Cells, Cultured
PubMed: 18782948
DOI: No ID Found -
Lasers in Surgery and Medicine Sep 2011A polyphenol constituent of green tea, epigallocatechin gallate (EGCG), has anti-carcinogenic properties. A growing number of studies document EGCG-mediated induction of...
BACKGROUND
A polyphenol constituent of green tea, epigallocatechin gallate (EGCG), has anti-carcinogenic properties. A growing number of studies document EGCG-mediated induction of apoptotic pathways and inhibition of pro-survival factors when combined with chemotherapy or radiation. We evaluated the efficacy of EGCG in modulating photofrin (PH)-mediated photodynamic therapy (PDT) responses.
METHODS
Mouse mammary carcinoma (BA) cells and transplanted BA tumors growing in C3H mice were treated with PH-mediated PDT. Select groups of treated cells and mice also received EGCG and then cytotoxicity, tumor response, and expression of survival molecules were evaluated in all experimental groups.
RESULTS
EGCG increased apoptosis and cytotoxicity in BA cells exposed to PH-mediated PDT. The initial pro-survival phase of the unfolded protein response (UPR), characterized by increased expression of the 78 kDa glucose-regulated protein (GRP-78), was induced by PDT. The second pro-apoptotic phase of the UPR, characterized by phospho-c-Jun N-terminal kinase (p-JNK) expression, activation of caspases-3 and 7, poly ADP ribose polymerase (PARP) cleavage, and expression of C/EBP homologous protein was observed when PDT was combined with EGCG. EGCG also decreased the expression of the pro-survival proteins GRP-78 and survivin, and attenuated PDT-induced prostaglandin E2 (PGE2 ) expression in PDT-treated cells. Comparable responses also were observed when BA tumors were treated with PDT and EGCG. In addition, PDT-induced expression of metalloproteinases (MMPs) and vascular endothelial growth factor (VEGF) was down-regulated in treated tumor tissue by EGCG.
CONCLUSIONS
The polyphenol EGCG improves PDT efficacy by increasing tumor apoptosis and decreasing expression of pro-survival and angiogenic molecules within the tumor microenvironment.
Topics: Animals; Antineoplastic Agents; Antineoplastic Combined Chemotherapy Protocols; Apoptosis; Blotting, Western; Catechin; Cell Line, Tumor; Dihematoporphyrin Ether; Dinoprostone; Electrophoresis, Polyacrylamide Gel; Endoplasmic Reticulum Chaperone BiP; Female; Heat-Shock Proteins; Inflammation; Mammary Neoplasms, Experimental; Matrix Metalloproteinases, Secreted; Mice; Mice, Inbred C3H; Photochemotherapy; Photosensitizing Agents; Unfolded Protein Response; Vascular Endothelial Growth Factors
PubMed: 22057492
DOI: 10.1002/lsm.21081 -
PloS One 2013Glioblastoma is the most common malignant brain tumor in humans. We explored the molecular mechanisms how the efficacy of photofrin based photodynamic therapy (PDT) was...
Photofrin based photodynamic therapy and miR-99a transfection inhibited FGFR3 and PI3K/Akt signaling mechanisms to control growth of human glioblastoma In vitro and in vivo.
Glioblastoma is the most common malignant brain tumor in humans. We explored the molecular mechanisms how the efficacy of photofrin based photodynamic therapy (PDT) was enhanced by miR-99a transfection in human glioblastoma cells. Our results showed almost similar uptake of photofrin after 24 h in different glioblastoma cells, but p53 wild-type cells were more sensitive to radiation and photofrin doses than p53 mutant cells. Photofrin based PDT induced apoptosis, inhibited cell invasion, prevented angiogenic network formation, and promoted DNA fragmentation and laddering in U87MG and U118MG cells harvoring p53 wild-type. Western blotting showed that photofrin based PDT was efficient to block the angiogenesis and cell survival pathways. Further, photofrin based PDT followed by miR-99a transfection dramatically increased miR-99a expression and also increased apoptosis in glioblastoma cell cultures and drastically reduced tumor growth in athymic nude mice, due to down regulation of fibroblast growth factor receptor 3 (FGFR3) and PI3K/Akt signaling mechanisms leading to inhibition of cell proliferation and induction of molecular mechanisms of apoptosis. Therefore, our results indicated that the anti-tumor effects of photofrin based PDT was strongly augmented by miR-99a overexpression and this novel combination therapeutic strategy could be used for controlling growth of human p53 wild-type glioblastomas both in vitro and in vivo.
Topics: Apoptosis; Blotting, Western; Cell Line, Tumor; Comet Assay; Dihematoporphyrin Ether; Flow Cytometry; Glioblastoma; Humans; In Vitro Techniques; MicroRNAs; Phosphatidylinositol 3-Kinases; Phosphoinositide-3 Kinase Inhibitors; Photochemotherapy; Proto-Oncogene Proteins c-akt; Real-Time Polymerase Chain Reaction; Receptor, Fibroblast Growth Factor, Type 3; Signal Transduction; Transfection
PubMed: 23409016
DOI: 10.1371/journal.pone.0055652 -
Cancer Biology & Therapy Dec 2012Patients with serosal (pleural or peritoneal) spread of malignancy have few definitive treatment options and consequently have a very poor prognosis. We have previously...
Patients with serosal (pleural or peritoneal) spread of malignancy have few definitive treatment options and consequently have a very poor prognosis. We have previously shown that photodynamic therapy (PDT) can be an effective treatment for these patients, but that the therapeutic index is relatively narrow. Here, we test the hypothesis that EGFR and STAT3 activation increase survival following PDT, and that inhibiting these pathways leads to increased PDT-mediated direct cellular cytotoxicity by examining BPD-PDT in OvCa and NSCLC cells. We found that BPD-mediated PDT stimulated EGFR tyrosine phosphorylation and nuclear translocation, and that EGFR inhibition by erlotinib resulted in reduction of PDT-mediated EGFR activation and nuclear translocation. Nuclear translocation and PDT-mediated activation of EGFR were also observed in response to BPD-mediated PDT in multiple cell lines, including OvCa, NSCLC and head and neck cancer cells, and was observed to occur in response to porfimer sodium-mediated PDT. In addition, we found that PDT stimulates nuclear translocation of STAT3 and STAT3/EGFR association and that inhibiting STAT3 signaling prior to PDT leads to increased PDT cytotoxicity. Finally, we found that inhibition of EGFR signaling leads to increased PDT cytotoxicity through a mechanism that involves increased apoptotic cell death. Taken together, these results demonstrate that PDT stimulates the nuclear accumulation of both EGFR and STAT3 and that targeting these survival pathways is a potentially promising strategy that could be adapted for clinical trials of PDT for patients with serosal spread of malignancy.
Topics: Antineoplastic Agents; Apoptosis; Cell Line, Tumor; Dihematoporphyrin Ether; ErbB Receptors; Erlotinib Hydrochloride; Female; Head and Neck Neoplasms; Humans; Lung Neoplasms; Ovarian Neoplasms; Photochemotherapy; Quinazolines; RNA Interference; RNA, Small Interfering; STAT3 Transcription Factor; Signal Transduction
PubMed: 22986230
DOI: 10.4161/cbt.22256 -
Blood Aug 1987To assess the potential of photoradiation therapy for the in vitro purging of residual tumor cells from autologous bone marrow (BM) transplants, we studied normal marrow...
To assess the potential of photoradiation therapy for the in vitro purging of residual tumor cells from autologous bone marrow (BM) transplants, we studied normal marrow and tumor cell clonogenicity in response to different light-activated compounds by using the fluorescent dyes dihematoporphyrin ether (DHE) and merocyanine-540 (MC-540). After photoradiation of cells with white light, both DHE and MC-540 showed high cytocidal activity toward lymphoid and myeloid neoplastic cells but had a significantly lesser effect on normal granulocyte-macrophage (CFU-GM), erythroid (BFU-E), and mixed colony-forming (CFU-GEMM) progenitor cells. Acute promyelocytic leukemia (HL-60), non-B, non-T, CALLA-positive acute lymphoblastic leukemia (Reh), and diffuse histocytic B cell lymphoma (SK-DHL-2) cell lines were exposed to different drug concentrations in combination with white light at a constant illumination rate of 50,000 lux. With DHE doses varying from 2.0 to 2.5 micrograms/mL and MC-540 concentrations of 15 to 20 micrograms/mL, clonogenic tumor cells could be reduced by more than 4 logs when treated alone or in mixtures with normal irradiated human marrow cells. However, preferential cytotoxicity towards neoplastic cells was highly dependent on the mode of light activation. MC-540 had no substantial effect on malignant lymphoid (SK-DHL-2) and myeloid (HL-60) cells and on normal marrow myeloid (CFU-GM) precursors when drug incubation was performed in the dark and followed by light exposure of washed cells. Equal doses of MC-540 (15 to 20 micrograms/mL) could preferentially eliminate tumor cells under conditions of simultaneous light and drug treatment (30 minutes at 37 degrees C). When using DHE (2.5 micrograms/mL), 29.3%, 46.8%, and 27.5% of normal marrow CFU-GM, BFU-E, and CFU-GEMM, respectively, were spared after sequential drug and light exposure of cells, whereas simultaneous treatment reduced both normal (CFU-GM) and neoplastic cells below the limits of detection. In summary, our results indicate the usefulness of various photoradiation models for the ex vivo treatment of leukemic and lymphomatous bone marrow autografts.
Topics: Bone Marrow; Bone Marrow Cells; Bone Marrow Transplantation; Cell Line; Dihematoporphyrin Ether; Hematopoietic Stem Cells; Hematoporphyrins; Humans; Leukemia; Lymphoma; Phototherapy; Pyrimidinones; Radiation-Sensitizing Agents; Transplantation, Autologous
PubMed: 2955818
DOI: No ID Found -
Photochemistry and Photobiology Mar 2023Photodynamic therapy (PDT) has been used to treat malignant pleural mesothelioma. Current practice involves delivering light to a prescribed light fluence with a point...
Photodynamic therapy (PDT) has been used to treat malignant pleural mesothelioma. Current practice involves delivering light to a prescribed light fluence with a point source, monitored by eight isotropic detectors inside the pleural cavity. An infrared (IR) navigation system was used to track the location of the point source throughout the treatment. The recorded data were used to reconstruct the pleural cavity and calculate the light fluence to the whole cavity. An automatic algorithm was developed recently to calculate the detector positions based on recorded data within an hour. This algorithm was applied to patient case studies and the calculated results were compared to the measured positions, with an average difference of 2.5 cm. Calculated light fluence at calculated positions were compared to measured values. The differences between the calculated and measured light fluence were within 14% for all cases, with a fixed scattering constant and a dual correction method. Fluence-surface histogram (FSH) was calculated for photofrin-mediated PDT to be able to cover 80% of pleural surface area to 50 J cm (83.3% of 60 J cm ). The study demonstrates that it will be possible to eliminate the manual measurement of the detector positions, reducing the patient's time under anesthesia.
Topics: Humans; Photochemotherapy; Mesothelioma; Mesothelioma, Malignant; Dihematoporphyrin Ether; Algorithms
PubMed: 35996976
DOI: 10.1111/php.13697 -
British Journal of Cancer Apr 1989Spheroids derived from the human colon adenocarcinoma cell line, WiDr, were exposed to 10 micrograms ml-1 Photofrin II and irradiated with light (700 nm, 50 mW cm-2)....
Spheroids derived from the human colon adenocarcinoma cell line, WiDr, were exposed to 10 micrograms ml-1 Photofrin II and irradiated with light (700 nm, 50 mW cm-2). Compared with exponentially growing monolayer cultures, cells in spheroids of 100, 250 and 500 microns diameter were respectively 1.8, 2.5 and 22-fold less sensitive. The small resistance of plateau-phase cultures (1.3-fold) was insufficient to account for this marked spheroid size-dependent resistance. For monolayer cultures and for spheroids of 100 and 250 microns diameter, the results were the same whether irradiations were carried out pre- or post-trypsinisation. However, there was a difference for the largest spheroid size: when irradiations were carried out pre-trypsinisation, spheroids were more resistant than when irradiations were given post-trypsinisation. Drug extraction studies showed that there was no difference in the average drug uptake between cultures of exponentially growing or plateau-phase cells, and 100 microns diameter spheroids while 250 and 500 microns diameter spheroids took up proportionally 0.5 and 0.4 as much drug. Cell contact effects, drug heterogeneity between cells, hypoxia and problems in drug penetration are suggested as possible reasons for the resistance of large spheroids to photodynamic treatment.
Topics: Adenocarcinoma; Cell Survival; Colonic Neoplasms; Dihematoporphyrin Ether; Drug Resistance; Hematoporphyrin Photoradiation; Hematoporphyrins; Humans; Photochemotherapy; Tumor Cells, Cultured
PubMed: 2523722
DOI: 10.1038/bjc.1989.105 -
Advances in Clinical and Experimental... 2012Melanoma is the most severe of skin neoplasms as it may grow rapidly and metastasize. The application of photodynamic therapy (PDT) opens up new prospects in the...
BACKGROUND
Melanoma is the most severe of skin neoplasms as it may grow rapidly and metastasize. The application of photodynamic therapy (PDT) opens up new prospects in the treatment of this tumor. Numerous studies suggest that the exposure of tumor cells to PDT can lead to cellular and molecular mechanisms which mediate oxidative stress in cells.
OBJECTIVES
The aim of this study was to evaluate in vitro the influence of photodynamic therapy on the human melanoma Me45 cell line.
MATERIAL AND METHODS
Photofrin (Ph) was used as a photosensitizer.
RESULTS
Viability studies have shown that there are significant differences between cells after PDT and cells without irradiation. After 24 hours of incubation with a 20 microg/ml concentration of Ph and with irradiation, less than 20% of the cells survived. In the control (without PDT), 65% of the cells survived.
CONCLUSIONS
The mitochondrial localization of Ph is significant, as it may lead to disturbances of mitochondrial transmembrane potential and finally to apoptotic cell death. The expressions of manganese superoxide dismutase and heme oxygenase and the level of carbonyl and thiol groups are indicating factors for oxidative stress in Me45 cells.
Topics: Apoptosis; Cell Line, Tumor; Cell Survival; Dihematoporphyrin Ether; Heme Oxygenase-1; Humans; Melanoma; Mitochondria; Oxidative Stress; Photochemotherapy; Photosensitizing Agents; Protein Carbonylation; Skin Neoplasms; Superoxide Dismutase; Time Factors
PubMed: 23214281
DOI: No ID Found