ngr-peptide and Breast-Neoplasms

A promising direction in Biopharmaceuticals is the development of specific peptide-based systems to improve drug delivery. This approach may increase tumor specificity and drug penetration into the target cell. Similar systems have been designed for several antitumor drugs. However, for photodynamic therapy drugs, such studies are not yet enough. Previously, we have developed a method of inclusion of chlorin e6 (Ce6), a photosensitizer used in photodynamic therapy, in phospholipid nanoparticles with a diameter of up to 30 nm, and reported an increase in its effectiveness in the experiments in vivo. In this work, we propose to modify a previously developed delivery system for Ce6 by the addition of cell-penetrating (R7) and/or targeting NGR peptides. The interaction of the compositions developed with HepG2 and MCF-7 tumor cells is shown. The expression of CD13 protein with affinity to NGR on the surface of these cells has been studied using flow cytometry. The expression of this protein on the HepG2 cells and its absence on MCF-7 was demonstrated. After incubation of tumor cells with the resulting Ce6 compositions, we evaluated the cellular accumulation, photoinduced, and dark cytotoxicity of the drugs. After irradiation, the highest level of cytotoxicity was observed when R7 peptide was added to the system, either alone or in combination with NGR. In addition to R7, the NGR-motif peptide increased the internalization of Ce6 in HepG2 cells without affecting its photodynamic activity. In this work we also discuss possible mechanisms of action of the cell-penetrating peptide when attached to phospholipid nanoparticles.

Topics: Breast Neoplasms; Carcinoma, Hepatocellular; CD13 Antigens; Cell Survival; Cell-Penetrating Peptides; Chlorophyllides; Drug Carriers; Drug Compounding; Hep G2 Cells; Humans; Liver Neoplasms; MCF-7 Cells; Nanoparticles; Oligopeptides; Phospholipids; Photochemotherapy; Photosensitizing Agents; Porphyrins

Fibrinogen Asn-Gly-Arg motif can specifically recognize and bind to Aminopeptidase N (CD13) on vascular endothelial cells in newly formed tumor vessels. Adipose-derived stem cells can serve as ideal vectors for gene therapy because of their ability of migrating to tumor tissues. First, this study was aimed to design a new peptide (CNGRCLLII(KLAKLAK)2) named CNAK which contains cyclic Asn-Gly-Arg motif and test its biological activity against human umbilical vein endothelial cells. Second, we aimed to construct stably transfected adipose-derived stem cells which express the CNAK peptide and investigate their anti-angiogenic activity in vivo. Adipose-derived stem cells were employed to localize CNAK on vascular endothelial cells in tumors based on their homing property. First of all, the new peptide was synthesized, which effectively entered into CD13+ human umbilical vein endothelial cells and showed cytotoxicity against human umbilical vein endothelial cells. The peptide induced apoptosis of human umbilical vein endothelial cells in a time- and dose-dependent manner, inhibited the expression of Bcl-2, and promoted the expression of Caspase-3 in human umbilical vein endothelial cells. Furthermore, the migration and tube formation of human umbilical vein endothelial cells were inhibited by CNAK. Primary adipose-derived stem cells were then isolated and identified. Stably transfected adipose-derived stem cells which express CNAK peptide (CNAK-ASCs) were successfully established, and the migration of CNAK-ASCs was assessed. In vivo, CNAK-ASCs were found to inhibit the growth and angiogenesis of breast cancer xenografts. This effect may be through inhibiting the secretion of matrix metalloproteinase-2 and membrane type 1-matrix metalloproteinase in vivo. It was also found that CNAK-ASCs reduced the quantity of breast cancer stem cells in tumor tissues. Our data suggested that the new peptide CNAK containing Asn-Gly-Arg motif had anti-angiogenic activity in vitro and in vivo.

Topics: Adipose Tissue; Animals; Apoptosis; Breast Neoplasms; CD13 Antigens; Cell Line, Tumor; Female; Fibrinogen; Genetic Therapy; Human Umbilical Vein Endothelial Cells; Humans; Mice; Oligopeptides; Peptides; Stem Cells; Transfection; Xenograft Model Antitumor Assays

The aim of this study was to evaluate the efficiency of a targeted siRNA nano-delivery system to silence the expression of Bmi-1 and hTERT, and to verify the toxicity of this delivery system in MCF-7 breast cancer cells. The most effective Bmi-1 siRNA and hTERT siRNA sequences were selected using RT-PCR and Western blotting. The polyethyleneimine (PEI)/siRNA nano-condensate was synthesized using PEI and modified using an NGR peptide fragment for targeting to tumor cells. The vector morphology, particle size and zeta potential were observed using an atomic force microscope and a laser particle size analyzer. The MCF-7 breast cancer cell line was transfected with the vector, and cytotoxicity was tested by MTT assays. The transfection efficiency was evaluated by qRT-PCR and Western blotting. Changes in gene expression and apoptosis rate were measured by flow cytometry. The size of LPN carrier and the condensate particle was between 100 and 200 nm and the potentials were close to neutral. There was maximum transfection efficiency and no significant increase in toxicity at 15 pmol/L. Bmi-1 and hTERT expression decreased, but the inhibition rate increased in the hTERT siRNA group, the hTERT+Bmi-1 siRNA group and the hTERT+Bmi-1 siRNA group compared with the scrambled siRNA group and the control group. Moreover, the hTERT+Bmi-1 siRNA group had the highest level of gene silencing. The complex, composed of Lipo, PEI and siRNA, is low toxicity and efficient transfection vectors. The expression level of Bmi-1 and hTERT was decreased by the gene silencing of either Bmi-1 or hTERT, but the effects were more significant when both were silenced simultaneously.

Topics: Apoptosis; Breast Neoplasms; Cell Proliferation; Female; Gene Expression Regulation, Neoplastic; Humans; MCF-7 Cells; Nanomedicine; Nanoparticles; Oligopeptides; Polycomb Repressive Complex 1; Polyethyleneimine; RNA Interference; RNA, Small Interfering; RNAi Therapeutics; Telomerase; Time Factors; Transfection

In this study, a peptide-based dual-targeting magnetic resonance imaging (MRI) contrast agent (S8) was designed and synthesized. Arg-Gly-Asp (RGD) and Asn-Gly-Arg (NGR) were combined in the targeting vector so as to allow binding, on the surface of tumor cells, to integrin αvβ3 and aminopeptidase N (CD13), respectively. The longitudinal relaxivity (r1) value of S8 was 8.297 mM-1sec-1 at a magnetic field of 11.7 T, which is approximately double the r1 value (4.25 mM-1sec-1) of Magnevist, a commercially available contrast agent. MDA-MB-231 human breast cancer cells (which overexpress αvβ3) and human prostate cancer cells PC-3 (which overexpress CD13) were used to investigate the tumor‑targeting behavior of S8. The results from the present study indicate that the designed contrast agent, S8, targets both MDA-MB‑231 and PC-3 cells.

Topics: Breast Neoplasms; CD13 Antigens; Cell Line, Tumor; Contrast Media; Female; Humans; Integrin alphaVbeta3; Magnetic Resonance Imaging; Male; Oligopeptides; Prostatic Neoplasms

Fullerene (C60) has shown great potential in drug delivery. In this study we exploited modified fullerene (diadduct malonic acid-fullerene-Asn-Gly-Arg peptide [DMA-C60-NGR]) as an antitumor drug carrier in order to build a new tumor-targeting drug delivery system. We also investigated the synergistic enhancement of cancer therapy using photodynamic therapy (PDT) induced by DMA-C60-NGR and 2-methoxyestradiol (2ME). Cytotoxicity tests indicated that DMA-C60-NGR had no obvious toxicity, while our drug delivery system (DMA-C60-2ME-NGR) had a high inhibition effect on MCF-7 cells compared to free 2ME. The tumor-targeting drug delivery system could efficiently cross cell membranes, and illumination induced the generation of intracellular reactive oxygen species and DNA damage. Furthermore, DMA-C60-2ME-NGR with irradiation had the highest inhibition effect on MCF-7 cells compared to the other groups. DMA-C60-NGR combined with 2ME showed a good synergistic photosensitization effect for inhibiting the growth of MCF-7 cells, demonstrating that DMA-C60-2ME-NGR may be promising for high treatment efficacy with minimal side effects in future therapy.

Topics: 2-Methoxyestradiol; Animals; Apoptosis; Breast Neoplasms; Cell Proliferation; Cell Survival; DNA Fragmentation; Drug Carriers; Estradiol; Female; Fullerenes; Humans; Malonates; MCF-7 Cells; Mice; Oligopeptides; Photochemotherapy; Photosensitizing Agents; Reactive Oxygen Species

To establish a NIR (near infrared)-/pH-responsive and sustained-release tumor-targeting drug delivery system (SWNT-PEI/DOX/NGR).. Functionalized SWNTs with polymerised polymeric poly(ethylene imine) was linked NGR (Asn-Gly-Arg) tumor-targeting peptide by DSPE-PEG2000-Maleimide via the maleimide group and sulfhydryl group of cysteine, in the end, doxorubicin (DOX) was attached to SWNT-PEI to obtain a SWNT-PEI/DOX/NGR delivery system.. The SWNT-PEI/DOX/NGR delivery system has significantly sustained-release effect and the slow release of DOX in normal tissues contribute to reduced systemic toxicity, while under 808 nm NIR laser irradiation or under lower pH environment the release of DOX can be accelerated.. Due to hyperthermia sensitizer effect of DOX, chemo-photothermal exemplified by SWNT-PEI/DOX/NGR tumor-targeting delivery system is a promising approach to anticancer therapy in vivo or in vitro.

Topics: Animals; Antibiotics, Antineoplastic; Breast; Breast Neoplasms; Cell Line, Tumor; Delayed-Action Preparations; Doxorubicin; Drug Delivery Systems; Female; Humans; Hydrogen-Ion Concentration; Maleimides; Mice; Mice, Inbred BALB C; Mice, Nude; Nanotubes, Carbon; Oligopeptides; Phosphatidylethanolamines; Polyethylene Glycols