cyanine-dye-3 and Neoplasms

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

Topics: Animals; Antineoplastic Agents; Carbocyanines; Cathepsin B; Fluorescent Dyes; Fluorides; HeLa Cells; Humans; Lanthanoid Series Elements; Light; Metal Nanoparticles; Mice; Neoplasms; NIH 3T3 Cells; Photochemotherapy; Photosensitizing Agents; Reactive Oxygen Species; Rose Bengal; Xenograft Model Antitumor Assays; Yttrium

MicroRNAs (miRNAs), small noncoding endogenous RNA molecules, are emerging as promising biomarkers for early detection of various diseases and cancers. Practical screening tools and strategies to detect these small molecules are urgently needed to facilitate the translation of miRNA biomarkers into clinical practice. In this study, a label-free biosensing technique based on surface-enhanced Raman scattering (SERS), referred to as plasmonic coupling interference (PCI), was applied for the multiplex detection of miRNA biomarkers. The sensing mechanism of the PCI technique relies on the formation of a nanonetwork consisting of nanoparticles with Raman labels located between adjacent nanoparticles that are interconnected by DNA duplexes. Because of the plasmonic coupling effect of adjacent nanoparticles in the nanonetwork, the Raman labels exhibit intense SERS signals. Such effect can be modulated by the addition of miRNA targets of interest that act as inhibitors to interfere with the formation of this nanonetwork, resulting in a diminished SERS signal. In this study, the PCI technique is theoretically analyzed, and the multiplex capability for detection of multiple miRNA cancer biomarkers is demonstrated, establishing the great potential of PCI nanoprobes as a useful diagnostic tool for medical applications.

Topics: Biomarkers, Tumor; Carbocyanines; DNA Probes; Fluorescent Dyes; Humans; Metal Nanoparticles; MicroRNAs; Neoplasms; Rhodamines; RNA, Neoplasm; Sensitivity and Specificity; Silver; Spectrum Analysis, Raman; Surface Plasmon Resonance

γ-Tilmanocept ((99m)Tc-labeled-tilmanocept or [(99m)Tc]-tilmanocept) is the first mannose-containing, receptor-directed, radiolabeled tracer for the highly sensitive imaging of sentinel lymph nodes in solid tumor staging. To elucidate the mannose-binding receptor that retains tilmanocept in this microenvironment, human macrophages were used that have high expression of the C-type lectin mannose receptor (MR; CD206). Cy3-labeled tilmanocept exhibited high specificity binding to macrophages that was nearly abolished in competitive inhibition experiments. Furthermore, Cy3-tilmanocept binding was markedly reduced on macrophages deficient in the MR by small interfering RNA treatment and was increased on MR-transfected HEK 293 cells. Finally, confocal microscopy revealed colocalization of Cy3-tilmanocept with the macrophage membrane MR and binding of labeled tilmanocept to MR(+) cells (macrophages and/or dendritic cells) in human sentinel lymph node tissues. Together these data provide strong evidence that CD206 is a major binding receptor for γ-tilmanocept. Identification of CD206 as the γ-tilmanocept-binding receptor enables opportunities for designing receptor-targeted advanced imaging agents and therapeutics for cancer and other diseases.

Topics: Carbocyanines; Cells, Cultured; Dextrans; Flow Cytometry; HEK293 Cells; Humans; Immunohistochemistry; Lectins, C-Type; Lymph Nodes; Macrophages; Mannans; Mannose Receptor; Mannose-Binding Lectins; Microscopy, Confocal; Molecular Structure; Neoplasms; Protein Binding; Radiopharmaceuticals; Receptors, Cell Surface; RNA Interference; Sentinel Lymph Node Biopsy; Technetium Tc 99m Pentetate

A novel reduction and pH dual-sensitive nonviral vector for long-circulating and tumor-targeted siRNA delivery is described. The nanomedicine is negatively charged at neutral pH of bloodstream whereas it is positively charged at lower pH of tumor tissue (ca. 6.8). Interlayer crosslinking with disulfide bonds stabilizes the nanomedicine during blood circulation and allows quick intracellular siRNA release after endocytosis.

Topics: Animals; Carbocyanines; Cell Line, Tumor; Endocytosis; Genetic Vectors; Humans; Hydrogen-Ion Concentration; Mice; Mice, Nude; Microscopy, Confocal; Neoplasms; Optical Imaging; Oxidation-Reduction; Polyethylene Glycols; Polymers; RNA, Small Interfering; Spectroscopy, Fourier Transform Infrared; Transfection; Transplantation, Heterologous

A near-infrared fluorochrome, GPU-311, was designed, synthesized and evaluated for its application in non-invasive imaging of tumor hypoxia. Efficient synthesis was achieved by nucleophilic substitution and click chemistry ring using the bifunctional tetraethylene glycol linker 2 containing thiol and azide groups for the conjugation of the propargylated nitroimidazole 1 and the heptamethine cyanine dye 3 bearing a 2-chloro-1-cyclohexenyl ring. GPU-311 exhibited long excitation and emission wavelength (Ex/Em=785/802 nm) and a decent quantum yield (0.05). The water solubility and hydrophilicity of GPU-311 increased. After in vitro treatment of SUIT-2/HRE-Luc pancreatic cancer cells with GPU-311, a higher level of fluorescence was observed selectively in hypoxia than in normoxia. However, in vivo fluorescence imaging of a mouse xenograft model after GPU-311 administration revealed inadequate accumulation of GPU-311 in tumors due to its rapid elimination through the liver.

Topics: Animals; Carbocyanines; Cell Hypoxia; Cell Line, Tumor; Fluorescence; Fluorescent Dyes; Humans; Hydrophobic and Hydrophilic Interactions; Mice; Neoplasms; Nitroimidazoles; Solubility; Spectroscopy, Near-Infrared

Differential display (DD) is one of the most commonly used approaches for identifying differentially expressed genes. Despite the great impact of the method on biomedical research, there has been a lack of automation of DD technology to increase its throughput and accuracy for a systematic gene expression analysis. Most of previous DD work has taken a "shotgun" approach of identifying one gene at a time, with limited polymerase chain reaction (PCR) reactions set up manually, giving DD a low-technology and low-throughput image. With our newly created DD mathematical model, which has been validated by computer simulations, global analysis of gene expression by DD technology is no longer a shot in the dark. After identifying the "rate-limiting" factors that contribute to the "noise" level of DD method, we have optimized the DD process with a new platform that incorporates fluorescent digital readout and automated liquid handling. The resulting streamlined fluorescent DD (FDD) technology offers an unprecedented accuracy, sensitivity, and throughput in comprehensive and quantitative analysis of gene expression. We are using this newly integrated FDD technology to conduct a systematic and comprehensive screening for p53 tumor-suppressor gene targets.

Topics: Animals; Apoptosis; Blotting, Northern; Blotting, Western; Carbocyanines; Cloning, Molecular; Deoxyribonuclease I; DNA, Complementary; Fluorescent Dyes; Gene Expression Profiling; Gene Expression Regulation; Gene Expression Regulation, Neoplastic; Genes, p53; Humans; Models, Theoretical; Neoplasms; Polymerase Chain Reaction; RNA; RNA, Messenger; Sensitivity and Specificity; Software; Time Factors; Transcription, Genetic; Tumor Suppressor Protein p53

Expression microarrays have great potential for clinical use but variability of the results represents a challenge for reliable practical application. The amount of fluorescent dye used in microarray experiments is a significant source of variability that has not been systematically studied. Here we demonstrate that the quantity of Cy3 dye affects microarray results performed on tumor specimens. Signal-to-noise ratios and coefficients of variation are significantly improved by increasing Cy3 to 150-180 pmol, but any further increase does not improve the data. In conclusion, optimal amounts of dye reduce variability and improve reliability of expression microarray experiments.

Topics: Carbocyanines; Fluorescent Dyes; Gene Expression Profiling; Humans; Neoplasms; Oligonucleotide Array Sequence Analysis