ABSTRACT
It is highly desirable to develop a therapeutic, observable nanoparticle complex for specific targeting in cancer therapy. Growth hormone (GH) and its antagonists have been explored as cancer cell-targeting molecules for both imaging and therapeutic applications. In this study, a low toxicity, biocompatible, therapeutic, and observable GH-nanoparticle complex for specifically targeting growth hormone receptor (GHR) in cancer cells was synthesized by conjugating GH with green fluorescence protein and carboxylated nanodiamond. Moreover, we have shown that this complex can be triggered by laser irradiation to create a "nanoblast" and induce cell death in the A549 non-small-cell lung cancer cell line via the apoptotic pathway. This laser-mediated, cancer-targeting platform can be widely used in cancer therapy.
Subject(s)
Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacology , Biocompatible Materials/pharmacology , Growth Hormone/chemistry , Nanodiamonds/chemistry , Neoplasms/drug therapy , Receptors, Somatotropin/antagonists & inhibitors , Antineoplastic Agents/chemical synthesis , Biocompatible Materials/chemical synthesis , Biocompatible Materials/chemistry , Cell Death/drug effects , Cell Line, Tumor , Cell Proliferation/drug effects , Dose-Response Relationship, Drug , Drug Screening Assays, Antitumor , Green Fluorescent Proteins/chemistry , Humans , Lasers , Molecular Structure , Neoplasms/pathology , Particle Size , Structure-Activity Relationship , Succinimides/chemistry , Surface PropertiesABSTRACT
Carcinoembryonic antigen-related cell adhesion molecule 6 (CEACAM6) is over-expressed in pancreatic cancer cells, and it is associated with the progression of pancreatic cancer. We tested a single domain antibody (sdAb) targeting CEACAM6, 2A3, which was isolated previously from a llama immune library, and an Fc conjugated version of this sdAb, to determine how they affect the pancreatic cancer cell line BxPC3. We also compared the effects of the antibodies to gemcitabine. Gemcitabine and 2A3 slowed down cancer cell proliferation. However, only 2A3 retarded cancer cell invasion, angiogenesis within the cancer mass and BxPC3 cell MMP-9 activity, three features important for tumour growth and metastasis. The IC50s for 2A3, 2A3-Fc and gemcitabine were determined as 6.5µM, 8µM and 12nM, respectively. While the 2A3 antibody inhibited MMP-9 activity by 33% compared to non-treated control cells, gemcitabine failed to inhibit MMP-9 activity. Moreover, 2A3 and 2A3-Fc inhibited invasion of BxPC3 by 73% compared to non-treated cells. When conditioned media that were produced using 2A3- or 2A3-Fc-treated BxPC3 cells were used in a capillary formation assay, the capillary length was reduced by 21% and 49%, respectively. Therefore 2A3 is an ideal candidate for treating tumours that over-express CEACAM6.
Subject(s)
Antigens, CD/immunology , Carcinoma, Pancreatic Ductal/pathology , Cell Adhesion Molecules/immunology , Cell Movement/drug effects , Cell Proliferation/drug effects , Neovascularization, Pathologic/prevention & control , Pancreatic Neoplasms/pathology , Single-Domain Antibodies/pharmacology , Animals , Camelids, New World , Carcinoma, Pancreatic Ductal/blood supply , Cells, Cultured , Drug Evaluation, Preclinical , GPI-Linked Proteins/immunology , Human Umbilical Vein Endothelial Cells/drug effects , Human Umbilical Vein Endothelial Cells/physiology , Humans , Neoplasm Invasiveness , Neovascularization, Pathologic/pathology , Neovascularization, Physiologic/drug effects , Pancreatic Neoplasms/blood supply , Pancreatic NeoplasmsABSTRACT
Haptoglobin (Hp) is an acute phase protein that binds free hemoglobin (Hb), preventing Hb-induced oxidative damage in the vascular system. There are three phenotypes in human Hp, whose heterogeneous polymorphic structures and varying concentrations in plasma have been attributed to the cause of diseases and outcome of clinical treatments. Different phenotypes of Hp may be composed of the same subunits but different copy numbers, rendering their determination difficult by a single procedure. In this study, we have developed a simple, fast, reliable and sensitive method, using label-free nanogold-modified bioprobes coupled with self-development electrochemical impedance spectroscopy (EIS). By this method, probe surface charge transfer resistance is detected. The relative charge transfer resistance ratios for Hp 1-1, Hp 2-1 and Hp 2-2 were characterized. We were able to determine protein size difference within 3 nm, and the linear region of the calibration curve for Hp levels in the range of 90 pg ml(-1) and 90 µg ml(-1) (â¼1 fM to 1 pM). We surmise that similar approaches can be used to investigate protein polymorphism and altered protein-protein interaction associated with diseases.
Subject(s)
Dielectric Spectroscopy/methods , Gold/chemistry , Haptoglobins/analysis , Metal Nanoparticles/chemistry , Antibodies/metabolism , Antibody Specificity/immunology , Antigens/metabolism , Electrodes , Enzyme-Linked Immunosorbent Assay , Humans , Phenotype , Protein Binding , Protein Stability , Reproducibility of Results , Time FactorsABSTRACT
The mechanism underlying DNA charge transport is intriguing. However, poor conductivity of DNA makes it difficult to detect DNA charge transport. Metallic DNA (M-DNA) has better conducting properties than native DNA. Ni(2+) may chelate in DNA and thus enhance DNA conductivity. On the basis of this finding, it is possible to reveal the mechanisms underlying DNA charge transport. The conductivity of various Ni-DNA species such as single-stranded, full complement, or mismatched sequence molecules was systematically tested with ultraviolet absorption and electrical or chemical methods. The results showed that the conductivity of single-stranded Ni-DNA (Ni-ssDNA) was similar to that of a native DNA duplex. Moreover, the resistance of Ni-DNA with a single basepair mismatch was significantly higher than that of fully complementary Ni-DNA duplexes. The resistance also increased exponentially as the number of mismatched basepairs increased linearly after the tunneling current behavior predicted by the Simmons model. In conclusion, the charges in Ni(2+)-doped DNA are transported through the Ni(2+)-mediated π-π stacking corridor. Furthermore, Ni-DNA acts as a conducting wire and exhibits a tunneling barrier when basepair mismatches occur. This property may be useful in detecting single basepair mismatches.
Subject(s)
DNA/chemistry , Nickel/chemistry , Static Electricity , Base Pair Mismatch , Base Pairing , Base Sequence , DNA/genetics , Dielectric Spectroscopy , Electric Conductivity , Electrochemical Techniques , Electrodes , Electrons , Gold/chemistry , Molecular Sequence Data , Spectrophotometry, UltravioletABSTRACT
Rhythmic oscillations of up to 600 Hz in grouped neurons frequently occur in the brains of animals. These high-frequency oscillations can be sustained in calcium-free conditions and may be blocked by gap junction blockers, implying a key role for electrical synapses in oscillation generation. Mathematical theories have been developed to demonstrate oscillations mediated by electrical synapses without chemical modulation; however, these models have not been verified in animals. Here we report that oscillations of up to 686 Hz are induced by paired spikes of short spike intervals (SIs) in a junction-coupled network. To initiate oscillations, it was essential that the second spike was elicited during the relative refractory period. The second spike suffered from slow propagation speed and failure to transmit through a low-conductance junction. Thus, at the spike initiation site, paired spikes of short SIs triggered one transjunctional spike in the postsynaptic neuron. At distant synaptic sites, two transjunctional spikes were produced as the SI increased during spike propagation. Consequently, spike collision of these asymmetrical transjunctional spikes occurred in the interconnected network. The remaining single spike reverberated in a network serving as an oscillator center. Paired-spike-induced oscillations were modeled by computer simulation and verified electrophysiologically in a network that mediates the tail-flip escape response of crayfish.
