Purpose: Legumain, a novel asparaginyl endopeptidase, has been observed to be highly expressed in several types of tumors including colorectal cancer. However, there is no study examining the relationship of legumain expression to clinocopathologic and biological variables in colorectal cancers.Experimental Design: We investigated legumain expression in 164 primary colorectal cancers, 34 corresponding distant normal mucosa samples, 89 adjacent normal mucosa samples, and 33 lymph node metastases using immunohistochemistry. We also did Western blotting analysis on three additional colorectal cancers and three colonic cell lines.Results: Legumain expression was increased in primary tumors compared with distant or adjacent normal mucosa (P < 0.05), but there was no significant change between primary tumors and metastases (P > 0.05). Legumain expression was positively related to poorer differentiation/ mucinous carcinoma (P = 0.04), higher degree of necrosis (P = 0.03) and apoptosis (P < 0.0001), positive proliferating cell nuclear antigen (P < 0.0001) and p53 expression (P = 0.049), and had a positive tendency towards stromelysin 3 (P = 0.058) and PINCH positivity (P = 0.05). The patients with tumors that showed both weak and lower percentage of the legumain expression, either in tumor (P = 0.01) or in stroma (P = 0.04), had a better prognosis.Conclusions: The legumain expression may be involved in colorectal cancer development and have a prognostic value in the patients.

Achieving selective and sensitive carbohydrate-protein interactions (CPIs) using nanotechnology is an intriguing area of research. Here we demonstrate that the different shapes of gold nanoparticles (AuNPs) functionalized with monosaccharides tune the bacterial aggregations. The mechanism of aggregation revealed that the large number of surface interactions of rod shaped mannose-AuNPs with E. coli ORN 178 compared with spherical and star-shaped AuNPs exhibited higher avidity and sensitivity. Moreover, such sensitive binding can be used for effective inhibition of bacterial infection of cells.

Advances in shape-dependent nanoparticle (NP) research have prompted a close scrutiny of the behaviour of nanostructures in vitro and in vivo. Data pertaining to cellular uptake and site specific sequestration of different shapes of NPs will undoubtedly assist researchers to design better nano-probes for therapeutic and imaging purposes. Herein, we investigated the shape dependent uptake of glyco-gold nanoparticles (G-AuNPs) in different cancer cell lines. Specifically, we have compared the behaviour of spherical, rod and star AuNPs with mannose and galactose conjugations. In vitro experiments showed that the rod-AuNPs exhibited the highest uptake over that of the star and spherical counterparts. Further, an investigation of the mechanism of the uptake clearly demonstrated clathrin mediated endocytosis of the specific G-AuNPs. These results reveal the benefits of different G-AuNP shapes in carbohydrate-mediated interactions.

Glyconanotechnology offers a broad range of applications across basic and translation research. Despite the tremendous progress in glyco-nanomaterials, there is still a huge gap between the basic research and therapeutic applications of these molecules. It has been reported that complexity and the synthetic challenges in glycans synthesis, the cost of the high order in vivo models and large amount of sample consumptions limited the effort to translate the glyco-nanomaterials into clinical applications. In this regards, several promising simple animal models for preliminary, quick analysis of the nanomaterials activities has been proposed. Herein, we have studied a systematic evaluation of the toxicity, biodistribution of fluorescently tagged PEG and mannose-capped gold nanoparticles (AuNPs) of three different shapes (sphere, rod, and star) in the adult zebrafish model, which could accelerate and provide preliminary results for further experiments in the higher order animal system. ICP-MS analysis and confocal images of various zebrafish organs revealed that rod-AuNPs exhibited the fast uptake. While, star-AuNPs displayed prolong sequestration, demonstrating its potential therapeutic efficacy in drug delivery.

Perfluoro undecanoyl chain conjugated peptide nucleic acids (PNAs) show 2.5 to 3 fold higher cellular uptake efficiency in NIH 3T3 and HeLa cells compared to simple undecanoyl PNAs. Fluorination of PNAs leads to the formation of lower size (∼100-250 nm) nanoparticles compared to larger size (∼500 nm) nanoparticles from non-fluorinated PNAs, thereby improving the efficiency of cell penetration.

We have developed peptidomimetic sialic acid (Sia) biosensors using boronic acid and arginine groups on the peptide backbone. The designed peptides were conjugated to fluorescent streptavidin via biotin enabling the optical labeling of cells. This approach provides unique opportunities to detect Sia composition on the cell surfaces and filopodia.

BackgroundRadiotherapy is a well-established anti-cancer treatment. Although radiotherapy has been shown to significantly decrease the local relapse in rectal cancer patients, the rate of distant metastasis is still very high. The aim of this study was to evaluate whether AEG-1 is involved in radiation-enhanced migration and invasion in vitro and in a novel in vivo zebrafish model.ResultsMigration and invasion were decreased in all the AEG-1 knockdown cell lines. Furthermore, we observed that radiation enhanced migration and invasion, while AEG-1 knockdown abolished this effect. The results from the zebrafish embryo model confirmed the results obtained in vitro. MMP-9 secretion and expression were decreased in AEG-1 knockdown cells.Materials and MethodsWe evaluated the involvement of AEG-1 in migration and invasion and, radiation-enhanced migration and invasion by Boyden chamber assay in three colon cancer cell lines and respective stable AEG-1 knockdown cell lines. Furthermore, we injected those cells into zebrafish embryos and evaluated the amount of disseminated cells into the tail.ConclusionAEG-1 knockdown inhibits migration and invasion, as well as radiation-enhanced invasion both in vitro and in vivo. We speculate that this is done via the downregulation of the intrinsic or radiation-enhanced MMP-9 expression by AEG-1 in the cancer cells. This study also shows, for the first time, that the zebrafish is a great model to study the early events in radiation-enhanced invasion.

The constructs and study of combinatorial libraries of structurally defined homologous extracellular matrix (ECM) glycopeptides can significantly accelerate the identification of cell surface markers involved in a variety of physiological and pathological processes. Herein, we present a simple and reliable host-guest approach to design a high-throughput glyco-collagen library to modulate the primary and secondary cell line migration process. 4-Amidoadamantyl-substituted collagen peptides and β-cyclodextrin appended with mono- or disaccharides were used to construct self-assembled glyco-collagen conjugates (GCCs), which were found to be thermally stable, with triple-helix structures and nanoneedles-like morphologies that altered cell migration processes. We also investigated the glycopeptide's mechanisms of action, which included interactions with integrins and cell signaling kinases. Finally, we report murine wound models to demonstrate the real-time application of GCCs. As a result of our observations, we claim that the host-guest model of ECM glycopeptides offers an effective tool to expedite identification of specific glycopeptides to manipulate cell morphogenesis, cell differentiation metastatic processes, and their biomedical applications.