The ras-related protein RAB22A interacts with hypoxia-inducible factor 1-alpha (HIF-1α) in MDA-MB-231 breast cancer cells in hypoxia.

BACKGROUND: Recent studies suggest that hypoxia-inducible factor 1-alpha (HIF-1α) and the small GTPase protein Ras-related protein Rab-22 A (RAB22A) may be colocalized in the cytoplasm and that as a conequence they may enhance the formation of microvesicles in breast cancer cells under hypoxia. Therefore, we sought to determine whether these two proteins are present in intracellular complexes in breast carcinoma cells. METHODS AND RESULTS: Evaluation using molecular docking indicated that HIF-1α and RAB22A interact with each other. Co-immunoprecipitation of endogenous or ectopically expressed HIF-1α and RAB22A proteins in MDA-MB-231 breast cancer cells or HEK-293T cells demonstrated that endogenous HIF-1α and RAB22A can form an intracellular complex; however, transiently expressed HIF-1α and RAB22A failed to interact. Investigating RAB22A and HIF-1α interactions in various cancer cell lines under hypoxia may shed light on their roles in cancer cell survival and progression through regulation of intracellular trafficking by HIF-1α under hypoxic conditions. CONCLUSIONS: Our study is the first to reveal the potential involvement of HIF-1α in intracellular trafficking through physical interactions with the small GTPase protein RAB22A. We discuss the implications of our work on the role of exosomes and microvesicles in tumor invasiveness.

The cytotoxicity effect of a bis-MPA-based dendron, a bis-MPA-PEG dendrimer and a magnetite nanoparticle on stimulated and non-stimulated human blood lymphocytes.

Dendrimers and dendrons offer a high surface area and nanoscale size and magnetic nanoparticles can be easily detected and manipulated due to their magnetic properties. The aim of the present study is to investigate the in vitro toxicity of Polyester-8-hydroxyl-1-carboxyl bis-MPA dendron, generation 3 (bis-MPA), Hyperbranched G4-PEG6k-OH (PEG) dendrimer and magnetite nanoparticle (Fe3O4), in human lymphocytes. Cell viability assays were performed on non-stimulated and lipopolysaccharide (LPS) stimulated lymphocytes, after exposure to various concentrations of the nanoparticles, using the Trypan blue assay, Flow Cytometry with 7-Amino Actinomycin D fluorescent dye (7-AAD), as well as the 3-[4,5-dimethylthiazol-2-yl] 2,5 diphenyl tetrazolium bromide (MTT) colorimetric method. The results collectively showed that after 24 h both the dendron and dendrimer at 50 µM concentration exhibited low cytotoxicity to non-stimulated and stimulated lymphocytes. Magnetite nanoparticle (Fe3O4) in concentrations 50-1000 µg/mL revealed negligible cytotoxicity to stimulated and non-stimulated lymphocytes. Moreover, the amount of intercellular Reactive Oxygen Species with or without treatment was assessed by means of the DCFH-DA to evaluate the presence of any oxidative stress. We propose herein simple cytotoxicity tests which indicate that these nanoparticles, after further studying, can serve as ideal drug carriers.