tetra(4-n-methylpyridyl)porphine and Neoplasms

Photodynamic therapy (PDT) as a clinical cancer treatment method has been used to treat carcinomas in different organs, and G-quadruplex-based DNA nanocompartments serving as the carriers of cationic porphyrin photosensitizers, especially combined with cell-targeting aptamers, is considered to offer new opportunities for future cancer treatment. However, the structural features of G-quadruplex/aptamer complexes suitable for the capsulation of photosensitizers and target cell recognition is unexplored so far. In this study, unimolecular (UM), bimolecular (BM) and tetramolecular (TM) G-quadruplex structures were used as the drug loading compartments and grafted onto tumor cell-targeting aptamer Sgc8, constructing several targeting drug delivery vehicles (T-GMVs). Besides the binding affinity of resulting DNA architectures for target cells and cell recognition specificity were explored in a comparative fashion, the drug loading capability and cancer therapy efficacy were evaluated using TMPyP4 as the model porphyrin-based drug. The experimental results show that only TM G-quadruplex structure is suitable to combine with Sgc8 for the development of drug delivery vehicle and the as-prepared T-GMV- TMPyP4 complexes display the desirable cancer therapy efficacy, holding the potential application in the future cancer therapy. More importantly, T-GMV- TMPyP4 is expected to lay the scientific groundwork for the successful development of G-quadruplex-based photosensitizer drug delivery carriers for the targeted cancer therapy.

Topics: Aptamers, Nucleotide; Cell Line, Tumor; Drug Carriers; Drug Screening Assays, Antitumor; G-Quadruplexes; Humans; Neoplasms; Photochemotherapy; Photosensitizing Agents; Porphyrins

Programmable DNA-based nanostructures (

Topics: Animals; Carbocyanines; DNA; Fluorescent Dyes; Humans; Hypoxia; Light; MCF-7 Cells; Mice; Mitochondria; Nanostructures; Neoplasms; Nitroreductases; Photochemotherapy; Photosensitizing Agents; Porphyrins; Singlet Oxygen

Currently, the nanocomposites based on silicon nanoparticles (SiNPs) are usually limited to a single therapeutic modality, and the design of the SiNPs nanohybrids with multi-modal synergistic therapeutic functions is still worth being explored to achieve more effective treatment. Herein, we used mesoporous silica nanoparticle (MSN) as a nanoplatform, SiNPs and the photosensitizer 5,10,15,20-tetrakis (1-methyl 4-pyridinio) porphyrin tetra (p-toluenesulfonate) (TMPyP) were first embedded in the MSN and was further modified with folic acid (FA) to obtain the mesoporous silica nanocomposite (MSN@SiNPs@TMPyP-FA) for targeted two-photon-excited fluorescence imaging-guided photodynamic therapy (PDT) and chemotherapy. The embedded TMPyP could generate singlet oxygen to perform PDT under light irradiation, meanwhile the anticancer drugs doxorubicin (DOX) could be loaded for chemotherapy. Moreover, due to the two-photon excited fluorescence of SiNPs, the nanocomposite successfully achieved targeted two-photon fluorescence cellular imaging at the near-infrared (NIR) laser excitation, which could effectively avoid the interference of biological auto-fluorescence. And in vitro cytotoxicity assays revealed that the synergistic therapy combining PDT and chemotherapy exhibited high therapeutic efficacy for cancer cells.

Topics: A549 Cells; Antineoplastic Agents; Doxorubicin; Drug Delivery Systems; Humans; MCF-7 Cells; Nanoparticles; Neoplasms; Optical Imaging; Photochemotherapy; Photosensitizing Agents; Porphyrins; Silicon; Silicon Dioxide; Theranostic Nanomedicine

Photodynamic therapy is a clinically approved procedure for the treatment of neoplastic and other non-malignant diseases. Meso-tetrakis(N-methyl-4-pyridyl)porphyrin (TMPyP) is a photosensitizing agent which has been used in many applications. However, the use of TMPyP topically is limited due to its hydrophilicity. To overcome this problem, TMPyP was loaded in ethosomes. Three ethosomal formulae (A), (B) and (C) were prepared and characterized. Preparation (A) was chosen to be used in the in vitro and in vivo study, having the greatest encapsulation efficiency, the smallest size and the highest cumulative release percentage. The results of in vitro permeation study revealed that the ethosomal TMPyP was superior to the drug in the free form with permeation flux (3.92 μg cm

Topics: Animals; Drug Delivery Systems; Drug Liberation; Drug Stability; Male; Mice; Microscopy, Electron, Transmission; Neoplasms; Particle Size; Photochemotherapy; Photosensitizing Agents; Porphyrins; Skin Absorption; Survival Analysis

Photodynamic therapy (PDT) is a useful tool against cancer and various other diseases. PDT is capable to induce different cell death mechanisms, due to the PDT evoked reactive oxygen species (ROS) production and is dose dependent. It is known that cytoskeleton is responsible for numerous cell functions, including cell division, maintenance of cell shape, their adhesion ability and movement. PDT initiated redistribution and subsequent disintegration of cytoskeletal components that precedes cell death. Here was present our results in HeLa and G361 cells subjected to sublethal PDT treatments using α,β,χ,δ porphyrin-Tetrakis (1-methylpyridinium-4-yl) p-Toluenesulfonate porphyrin (TMPyP). The photosensitizer (PS) induced transient increasing of mitotic index (MI) observable early after PDT, cell cycle arrest, microtubule (MTs) disorganization of interphase cells, aberrant mitosis and formation of rounded cells with partial loss of adherence. Some cells were partly resistant to PDT induced MTs disorganization. The differences between both cell lines to PDT response were described. This is the first evidence of TMPyP - PDT induced microtubule disorganization and the cell death mechanisms known as mitotic catastrophe and the first detail analysis of microtubule aberrations of mitotic and interphase cells in HeLa and G361 cell lines. New modification of techniques of protein immunolabeling was developed.

Topics: Cell Cycle Checkpoints; Cell Line, Tumor; Cell Survival; Cytoskeleton; HeLa Cells; Histones; Humans; Light; Microscopy, Fluorescence; Microtubules; Mitosis; Neoplasms; Photochemotherapy; Photosensitizing Agents; Porphyrins; Reactive Oxygen Species

Topics: Animals; Antineoplastic Agents; Cell Survival; Doxorubicin; Drug Carriers; Flow Cytometry; Folic Acid; HeLa Cells; Humans; Light; Magnetics; Mice; Nanoparticles; Neoplasms; Optical Imaging; Photosensitizing Agents; Porphyrins; RNA

Previous studies have shown that promoter regions of many proto-oncogenes can fold into G-quadruplexes, which are potentially involved in the regulation of genes. Bioinformatics analysis suggested that there was a G-rich sequence within -48 to -26 region of the human MET promoter (named Pu23WT). In this study, we proved that Pu23WT adopted an intramolecular parallel G-quadruplex structure under physiological conditions in vitro, and the cationic porphyrin TMPyP4 enhanced the stability of the Pu23WT G-quadruplex. To better understand the functions of Pu23WT in the MET expression, we performed a series of analysis on several cancer cells. Experimental data revealed that TMPyP4 treatment attenuated the expression of MET in HepG2, BGC823, and U87MG cells, resulting in the cellular proliferation inhibition, G1 phase cell cycle arrest and cell migration retardation. ChIP assay results indicated that TMPyP4 probably prohibited the Pu23WT G-quadruplex from binding to the activator Sp1, which could be one of the mechanisms that led to the transcription inhibition of MET gene. It is the first study on the G-quadruplex structure in the human MET promoter and its functions in cancer cells. We believe that this structure is a potential target for anticancer treatment.

Topics: Cell Line, Tumor; Cell Movement; Cell Proliferation; G-Quadruplexes; Hep G2 Cells; Humans; Neoplasms; Nucleic Acid Conformation; Porphyrins; Promoter Regions, Genetic; Proto-Oncogene Proteins c-met; Sp1 Transcription Factor

TMPyP4 is widely considered as a potential photosensitizer in photodynamic therapy and a G-quadruplex stabilizer for telomerase-based cancer therapeutics. However, its biological effects including a possible adverse-effect are poorly understood. In this study, whole genome RNA-seq analysis was used to explore the alteration in gene expression induced by TMPyP4. Unexpectedly, we find that 27.67% of changed genes were functionally related to cell adhesion. Experimental evidences from cell adhesion assay, scratch-wound and transwell assay indicate that TMPyP4 at conventional doses (≤0.5 μM) increases cell-matrix adhesion and promotes the migration of tumor cells. In contrast, a high dose of TMPyP4 (≥2 μM) inhibits cell proliferation and induces cell death. The unintended "side-effect" of TMPyP4 on promoting cell migration suggests that a relative high dose of TMPyP4 is preferred for therapeutic purpose. These findings contribute to better understanding of biological effects induced by TMPyP4 and provide a new insight into the complexity and implication for TMPyP4 based cancer therapy.

Topics: A549 Cells; Cell Death; Cell Movement; Cell Proliferation; Dose-Response Relationship, Drug; HeLa Cells; Humans; Neoplasms; Porphyrins

Photodynamic therapy (PDT) is based on the tumor-selective accumulation of photosensitizer followed by irradiation with light of an appropriate wavelength. After irradiation and in the presence of oxygen, photosensitizer induces cellular damage. The aim of this study was to evaluate effects of two photosensitizers TMPyP and ClAlPcS2 on cell lines to obtain better insight into their mechanisms of action. We determined cell viability, reactive oxygen species (ROS) generation and changes in expression levels of two important early response genes, C-MYC and C-FOS, on tumor MCF7 (human breast adenocarcinoma) and G361 (human melanoma) cell lines and non-tumor BJ cell line (human fibroblast) after photodynamic reaction with TMPyP and ClAlPcS2 as photosensitizers. In addition TMPyP and ClAlPcS2 cellular uptake and clearance and antioxidant capacity of the mentioned cell lines were investigated. We found appropriate therapeutic doses and confirmed that both tested photosensitizers are photodynamically efficient in treatment used cells in vitro. TMPyP is more efficient; it had higher ROS production and toxicity after irradiation by intermediate therapeutic doses than ClAlPcS2. We revealed that both TMPyP and ClAlPcS2-PDT increased C-FOS expression on tumor cell lines (G361 and MCF7), but not on non-tumor BJ cell line. Conversely, both TMPyP and ClAlPcS2-PDT decreased C-MYC expression on non-tumor BJ cell line but not on tumor cell lines. As first we tested these photosensitizers in such extent and we believe that it can help to better understand mechanisms of PDT and increase its efficiency and applicability.

Topics: Antioxidants; Cell Line, Tumor; Cell Survival; Humans; Indoles; Light; MCF-7 Cells; Neoplasms; Organometallic Compounds; Photochemotherapy; Photosensitizing Agents; Porphyrins; Proto-Oncogene Proteins c-fos; Proto-Oncogene Proteins c-myc; Reactive Oxygen Species; Up-Regulation

DNA and porphyrin based therapeutics are important for anti-cancer treatment. The present studies demonstrate single-stranded DNA (ssDNA) assembles with meso-tetra-4-pyridyl porphine (MTP) forming porphyrin:DNA nano-complexes (PDN) that are stable in aqueous solution under physiologically relevant conditions and undergo dissociation with DNA release in hydrophobic environments, including cell membranes. PDN formation is DNA-dependent with the ratio of porphyrin:DNA being approximately two DNA nucleobases per porphyrin. PDN produce reactive oxygen species (ROS) in a light-dependent manner under conditions that favor nano-complex dissociation in the presence of hydrophobic solvents. PDN induce light-dependent cytotoxicity in vitro and anti-tumor activity towards bladder cancer xenografts in vivo. Light-dependent, PDN-mediated cell death results from ROS-mediated localized membrane damage due to lipid peroxidation with mass spectrometry indicating the generation of the lipid peroxidation products 9- and 13-hydroxy octadecanoic acid. Our results demonstrate that PDN have properties useful for therapeutic applications, including cancer treatment.. In this study, porphyrin-DNA nanocomplexes were investigated as anti-cancer therapeutics inducing ROS production in a light-dependent manner. Efficacy is demonstrated in vitro as well as a in a bladder cancer xenograft model.

Topics: Animals; Antineoplastic Agents; Cell Death; Cell Line, Tumor; Cell Membrane; DNA, Single-Stranded; Endosomes; Female; Humans; Hydrophobic and Hydrophilic Interactions; Lipid Peroxidation; Mice; Mice, Nude; Nanomedicine; Neoplasm Transplantation; Neoplasms; Photosensitizing Agents; Porphyrins; Reactive Oxygen Species; Stearic Acids; Urinary Bladder Neoplasms

Meso-tetra-(N-methylpiridinium-4-yl)-porphyrin (TMPyP) and meso-tetra-(4-sulfonatophenyl)-porphyrin (TPPS(4)) are photosensitizing drugs (PS) used in photodynamic therapy (PDT). Based on the fact that these compounds present similar chemical structures but opposite charges at pH levels near physiological conditions, this work aims to evaluate the in vitro and in vivo influence of these electrical charges on the iontophoretic delivery of TMPyP and TPPS(4), attempting to achieve maximum accumulation of PS in skin tissue. The iontophoretic transport of these drugs from a hydrophilic gel was investigated in vitro using porcine ear skin and vertical, flow-through diffusion cells. In vivo experiments using rats were also carried out, and the penetration of the PSs was analyzed by fluorescence microscopy to visualize the manner of how these compounds were distributed in the skin after a short period of iontophoresis application. In vitro, both passive and iontophoretic delivery of the positively charged TMPyP were much greater (20-fold and 67-fold, respectively) than those of the negatively charged TPPS(4). TPPS(4) iontophoresis in vivo increased the fluorescence of the skin only in the very superficial layers. On the other hand, iontophoresis of the positively charged drug expressively increased the rat epidermis and dermis fluorescence, indicating high amounts of this drug throughout the skin layers. Moreover, TMPyP was homogeneously distributed around and into the nuclei of the skin cells, suggesting its potential use in topical PDT.

Topics: Administration, Cutaneous; Administration, Topical; Animals; Cellulose; Diffusion; Ear; Iontophoresis; Male; Neoplasms; Photochemotherapy; Porphyrins; Rats; Rats, Wistar; Skin; Skin Absorption; Swine

A specialized G-rich DNA structure, G-quadruplex, has been studied for its special physical characteristics and biological effects. Herein we report a novel strategy of using G-quadruplex as a drug carrier to target cancer cells for photodynamic therapy (PDT). A G-quadruplex forming AS1411 aptamer could be physically conjugated with six molecules of porphyrin derivative, 5,10,15,20-tetrakis(1-methylpyridinium-4-yl)porphyrin (TMPyP4), to fabricate the apt-TMP complex. The TMPyP4 molecules in the complex were identified to bind tightly to the aptamer by intercalation and outside binding. Because the G-quadruplex structure is known to target the overexpressed nucleolin in cancer cells, in this study, the effect of the G-quadruplex structure as a carrier for the delivery of TMPyP4 into cancer cells by nucleolin-mediated internalization was investigated. The results showed that the apt-TMP complex exhibited a higher TMPyP4 accumulation in MCF7 breast cancer cells than in M10 normal epithelium cells. After treated with light for 180 s, the photodamage in MCF7 cells was larger than in M10 cells. These results indicated that the TMPyP4 delivery and uptake were mediated by the specific interaction of the apt-TMP complex with nucleolin on the cellular surface and that the use of the AS1411 aptamer as a drug carrier may be a potential tactic in cancer therapy.

Topics: Aptamers, Nucleotide; Base Sequence; Biological Transport; Cell Line, Tumor; Cell Nucleus; Circular Dichroism; Drug Carriers; Flow Cytometry; G-Quadruplexes; Gene Expression Regulation, Neoplastic; Humans; Ligands; Microscopy, Electron, Transmission; Neoplasms; Nucleolin; Organ Specificity; Phosphoproteins; Photochemotherapy; Porphyrins; RNA-Binding Proteins; Spectrophotometry, Ultraviolet

A series of cationic porphyrins has been identified as G-quadruplex interactive agents (QIAs) that stabilize telomeric G-quadruplex DNA and thereby inhibit human telomerase; 50% inhibition of telomerase activity was achieved in HeLa cell-free extract at porphyrin concentrations in the range < or = 50 microM. Cytotoxicity of the porphyrins in vitro was assessed in normal human cells (fibroblast and breast) and human tumor cells representing models selected for high telomerase activity and short telomeres (breast carcinoma, prostate, and lymphoma). In general, the cytotoxicity (EC50, effective concentration for 50% inhibition of cell proliferation) against normal and tumor cells was > 50 microM. The porphyrins were readily absorbed into tumor cell nuclei in culture. Inhibition of telomerase activity in MCF7 cells by subcytotoxic concentrations of TMPyP4 showed time and concentration dependence at 1-100 microM TMPyP4 over 15 days in culture (10 population doubling times). The inhibition of telomerase activity was paralleled by a cell growth arrest in G2-M. These results suggest that relevant biological effects of porphyrins can be achieved at concentrations that do not have general cytotoxic effects on cells. Moreover, the data support the concept that a rational, structure-based approach is possible to design novel telomere-interactive agents with application to a selective and specific anticancer therapy.

Topics: Antineoplastic Agents; Breast Neoplasms; Cations; Cell Nucleus; DNA; DNA, Neoplasm; Enzyme Inhibitors; Female; Fibroblasts; G-Quadruplexes; HeLa Cells; Humans; Models, Molecular; Neoplasms; Porphyrins; Telomerase; Tumor Cells, Cultured