5-carboxytetramethylrhodamine-succinimidyl-ester and Neoplasms

Investigating the multimolecule patterns in living cells is of vital importance for clinical and biomedical studies. Herein, we reported for the first time the engineering of gold nanoflares as smart automata to implement computing-based diagnosis in living mammalian cells. Defining the logic combinations of miR122 and miR21 as the detection patterns, the corresponding OR and AND diagnostic automata were designed. The results showed that they could recognize the correct patterns rapidly and sensitively. The automata could enter cells via self-delivery and have good biocompatibility. They enabled accurate diagnosis on miRNA signatures in different cell lines and differentiation of fluctuations in the same cell line at single cell resolution. Moreover, the automata afforded an innovative diagnostic mode. It simplified the complicated process of detecting, data-collecting, computing, and evaluating. The direct diagnosing result ("1" or "0") was exported according to the embedded computation code. It highlighted the new possibility of using smart automata for intelligent diagnostics and cancer therapy at single cell resolution.

Topics: Cell Line, Tumor; Computers, Molecular; Fluorescent Dyes; Gold; HEK293 Cells; Humans; Logic; Metal Nanoparticles; MicroRNAs; Microscopy, Confocal; Microscopy, Fluorescence; Neoplasms; Nucleic Acid Hybridization; Oligonucleotides; Rhodamines

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

A novel tumor-selective catalytic nanosystem that enables efficient chemodynamic therapy (CDT) and activatable fluorescence imaging in H2O2-rich tumor microenvironments has been developed.

Topics: Antineoplastic Agents; Apoptosis; DNA Probes; Fluorescence; Fluorescent Dyes; Graphite; HeLa Cells; Hepatocytes; Humans; Hydrogen Peroxide; Metal Nanoparticles; Microscopy, Confocal; Microscopy, Fluorescence; Neoplasms; Oxides; Rhodamines; Silver; Theranostic Nanomedicine

Sentinel lymph nodes (SLN) are the first lymph nodes (LN) where cancer cells metastasize from the primary tumor. We designed fluorophore-quencher-based activatable nanoparticles for SLN imaging. We selected TAMRA as a fluorophore and BHQ2 or QSY7 as a quencher. Ternary anionic complexes were constructed with generation 4th polyamidoamine dendrimer (G4) modified with TAMRA and p-SCN-Bn-DTPA (DTPA), polyethyleneimine (PEI) modified with BHQ2 or QSY7, and γ-polyglutamic acid (γ-PGA) by the electrostatic self-assembly system. TAMRA-G4-DTPA/PEI-BHQ2/γ-PGA and TAMRA-G4-DTPA/PEI-QSY7/γ-PGA complexes had a particle size of about 40 nm and a ζ-potential of -50 mV, and showed fluorescence resonance energy transfer (FRET) quenching. Fluorescence microscopy studies demonstrated that TAMRA-G4-DTPA/PEI-QSY7/γ-PGA complex produced intracellular fluorescent signals in the lysosome. During in vivo fluorescent imaging, TAMRA-G4-DTPA/PEI-QSY7/γ-PGA complex enabled the detection of mouse popliteal LN. The fluorophore-quencher conjugated γ-PGA complex based on FRET quenching would be useful for fluorescence-based optical imaging of SLN.

Topics: Animals; Dendrimers; Fluorescence; Fluorescent Dyes; Humans; Lymphatic Metastasis; Lysosomes; Male; Mice, Inbred BALB C; Nanoparticles; Neoplasms; Optical Imaging; Particle Size; Pentetic Acid; Polyamines; Polyethyleneimine; Polyglutamic Acid; Rhodamines; Sentinel Lymph Node

The serine-aspartic acid-valine (SDV) peptide binds specifically to integrin α

Topics: Animals; Cell Line, Tumor; Contrast Media; HT29 Cells; Humans; Integrin alphaVbeta3; Mice; Multimodal Imaging; Neoplasms; Neovascularization, Pathologic; Peptides; Rhodamines; Technetium

PNC-27, a peptide that contains an HDM-2-binding domain from p53 attached to a membrane-penetrating peptide on its carboxyl terminal end, is cytotoxic to cancer, but not normal, cells. It forms transmembrane pores in the cancer cell membrane. Our purpose is to determine if the whole peptide or critical fragments induce pore formation in cancer cells.. We have prepared PNC-27 with a green fluorescent label on its amino terminus and a red fluorescent label on its carboxyl terminus and treated MCF-7 breast cancer cells and untransformed MCF-10-2A breast epithelial cells with this double-labeled peptide to determine if combined yellow fluorescence occurs in the membrane of the cancer cells during cancer cell killing.. At 30 min, there is significant combined punctate yellow fluorescence, indicative of intact peptide, in the cell membrane of cancer cells that increases during cancer cell lysis. MCF-10-2A cells show initial (30 min) uniform combined yellow membrane fluorescence that subsequently disappears. Unlike the cancer cells, these untransformed cells remain viable.. PNC-27 induces cancer cell membrane lysis by acting as the whole peptide, not fragments. The punctate yellow fluorescence is due to interaction of PNC-27 with intramembrane targets of MCF-7 cells that do not exist in the membrane of the untransformed cell line. This interaction increases the lifetime of PNC-27. Absence of these targets in the membranes of the untransformed MCF-10-2A cells results in initial uniform fluorescence of the double-labeled peptide in their membranes after which the peptide is degraded.

Topics: Antineoplastic Agents; Apoptosis; Caspases; Cell Line, Tumor; Cell Membrane; Coloring Agents; Fluorescein-5-isothiocyanate; Fluorescent Dyes; Green Fluorescent Proteins; Humans; L-Lactate Dehydrogenase; Microscopy, Confocal; Necrosis; Neoplasms; Rhodamines; Tetrazolium Salts; Thiazoles; Tumor Suppressor Protein p53