ABSTRACT
Multidrug resistance (MDR) refers to the ability of cancer cells to develop cross resistance to a range of anticancer drugs which are structurally and functionally unrelated. P-glycoprotein (P-gp) is the best studied MDR phenotype in photodynamic therapy (PDT) treated cells. Our pervious study demonstrated that FosPeg® mediated PDT is effective to NPC cell line models. In this in vitro study, the expression of MDR1 gene and its product P-gp in undifferentiated, poorly differentiated and well differentiated human nasopharyngeal carcinoma (NPC) cells were investigated. The influence of P-gp efflux activities on photosensitizer FosPeg® was also examined. Regardless of the differentiation status, PDT tested NPC cell lines all expressed P-gp protein. Results indicated that FosPeg® photoactivation could heighten the expression of MDR1 gene and P-gp transporter protein in a dose dependent manner. Up to 2-fold increase of P-gp protein expression were seen in NPC cells after FosPeg® mediated PDT. Interestingly, our finding demonstrated that FosPeg® mediated PDT efficiency is independent to the MDR1 gene and P-gp protein expression in NPC cells. FosPeg® itself is not the substrate of P-gp transporter protein and no efflux of FosPeg® were observed in NPC cells. Therefore, the PDT efficiency would not be affected even though FosPeg® mediated PDT could induce MDR1 gene and P-gp protein expression in NPC cells. FosPeg® mediated PDT could be a potential therapeutic approach for MDR cancer patients.
Subject(s)
Gene Expression Regulation/drug effects , Liposomes/pharmacology , Mesoporphyrins/pharmacology , Photosensitizing Agents/pharmacology , ATP Binding Cassette Transporter, Subfamily B/genetics , ATP Binding Cassette Transporter, Subfamily B/metabolism , Carcinoma , Cell Line, Tumor , Drug Resistance, Neoplasm/drug effects , Gene Expression Regulation/radiation effects , Humans , Light , Liposomes/chemistry , Liposomes/therapeutic use , Mesoporphyrins/chemistry , Mesoporphyrins/therapeutic use , Nasopharyngeal Carcinoma , Nasopharyngeal Neoplasms/drug therapy , Nasopharyngeal Neoplasms/metabolism , Nasopharyngeal Neoplasms/pathology , Photochemotherapy , Photosensitizing Agents/chemistry , Photosensitizing Agents/therapeutic use , RNA, Messenger/metabolismABSTRACT
Nasopharyngeal carcinoma (NPC) is one of the top ten cancers highly prevalent in Hong Kong and South China. Epstein-Barr virus (EBV) infection contributes to the tumorigenesis of NPC through the expression of different viral proteins. Among these, Latent Membrane Protein 1(LMP1) is the major oncoprotein expressed by EBV. Foscan® (Biolitec AG), m-tetrahydroxyphenylchlorin (mTHPC)-based photosensitizing drug, has been used in the photodynamic therapy (PDT) for head and neck cancers. FosPeg® (Biolitec AG) is a new formulation of mTHPC contained in PEGylated liposomes with optimized distribution properties. In this in vitro study, the potential of FosPeg®-PDT on human EBV positive NPC cell (c666-1) and EBV negative cells (HK1 and CNE2) were investigated. Effects of FosPeg®-PDT on the expression of EBV BART miRNAs (EBV miRNA BART 1-5p, BART 16, and BART 17-5p), LMP1 mRNA and proteins on c666-1 cells were also elucidated. The killing efficacy of FosPeg®-PDT on NPC cells were determined by MTT assay after LED activation. Effects of FosPeg®-PDT on the expression of LMP1 mRNA and protein were examined by real time PCR and western blot analysis. FosPeg®-PDT demonstrated its antitumor effect on c666-1 cells in a drug and light dose dependent manner. LD30, LD50 and LD70 were achieved by applying LED activation (3J/cm(2)) at 4h post incubated cells with 0.05µg/ml, 0.07µg/ml and 0.3µg/ml FosPeg®, respectively. Up-regulation of both LMP1 mRNA and protein were observed after FosPeg®-PDT in a dose dependent manner. FosPeg®-PDT exerted antitumor effect on c666-1 cells through up-regulation of LMP1 protein. Understanding the mechanism of FosPeg®-PDT may help to develop better strategies for the treatment of NPC.
Subject(s)
Gene Expression Regulation, Neoplastic/drug effects , Herpesvirus 4, Human/genetics , Mesoporphyrins/pharmacology , MicroRNAs/genetics , Nasopharyngeal Neoplasms/pathology , Photochemotherapy , Viral Matrix Proteins/metabolism , Cell Cycle/drug effects , Cell Cycle/radiation effects , Cell Line, Tumor , Chemistry, Pharmaceutical , Dose-Response Relationship, Drug , Gene Expression Regulation, Neoplastic/radiation effects , Herpesvirus 4, Human/physiology , Humans , Intracellular Space/drug effects , Intracellular Space/metabolism , Intracellular Space/radiation effects , Liposomes , Mesoporphyrins/administration & dosage , Mesoporphyrins/chemistry , Mesoporphyrins/metabolism , Mesoporphyrins/therapeutic use , Nasopharyngeal Neoplasms/virology , Polyethylene Glycols/chemistry , RNA, Viral/genetics , Viral Matrix Proteins/geneticsABSTRACT
Tissue damage in the CNS is critically influenced by the adaptive immune system. Primary oligodendrocyte damage (by overexpression of PLP) leads to low-grade inflammation of high pathological impact, which is mediated by CD8+ T cells. To yield further insight into pathogenesis and nature of immune responses in myelin mutated mice, we here apply a detailed immunological characterization of CD8+ T cells in PLP-transgenic and aged wild type mice. We provide evidence that T effector cells accumulate in the CNS of PLP-transgenic and wild-type mice and show a higher level of activation in mutant mice, indicated by surface markers and clonal expansions, as demonstrated by T cell receptor CDR3-spectratype analysis. Vbeta-Jbeta similarities suggest specificity against a common antigen, albeit we could not find specific responses against myelin-antigen-derived peptides. The association of primary oligodendrocyte damage with secondary expansions of pathogenic cells underlines the role of adaptive immune reactions in neurodegenerative and neuroinflammatory diseases.
Subject(s)
CD8-Positive T-Lymphocytes/immunology , Central Nervous System/immunology , Demyelinating Diseases/immunology , Immune System/immunology , Myelin Sheath/immunology , Animals , Autoimmunity/genetics , Autoimmunity/immunology , CD8 Antigens/immunology , Cell Proliferation , Central Nervous System/metabolism , Central Nervous System/physiopathology , Clone Cells/immunology , Demyelinating Diseases/genetics , Demyelinating Diseases/physiopathology , Immune System/physiopathology , Immunity, Innate/genetics , Immunity, Innate/immunology , Lymphocyte Activation/genetics , Lymphocyte Activation/immunology , Mice , Mice, Inbred C57BL , Mice, Neurologic Mutants , Mice, Transgenic , Myelin Sheath/genetics , Myelin Sheath/metabolism , Oligodendroglia/immunology , Oligodendroglia/metabolism , Oligodendroglia/pathology , T-Lymphocyte Subsets/immunology , T-Lymphocyte Subsets/metabolismABSTRACT
The clinical and pathological features of an apparently unique case of an endometrial cyst of the uterus are reported. The cyst was located within the myometrium of a 16-year-old woman suffering from dysmenorrhea. After excision of the cyst, patient's symptoms improved. On histological examination, the cyst most closely resembled an adenomyotic cyst.
