Molecular and morphological study of SDF-1 in tumor proliferation of pituitary adenoma

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In the present study, we aimed to develop a novel transfection method for short interference RNA (siRNA) to employ nanotube by the surfactant activity.A6K consisting of six alanine residues and hydrophilic head, lysine, was compared with conventional cationic transfectant reagent, siFECTOR and Lipofectamine 2000. Cyto-toxity for human rat pituitary adenoma cell line, AtT-20 or GH3 was examined with MTS assay. Transfection efficiency was analyzed with FITC-labelled siRNA targeting SDF-1 receptor mRNA with fluorescent activity and microscopy. Ultra-structure of A6K was observed with electron microscopy.Non-cytotoxic level of A6K (IC50, 320.8 ・g/ml) was significantly higher than that of siFECTOR ((IC50, 35.6 ・g/ml) and Lipofectamine 2000 (IC50, 22.4 ・g/ml). Transfection efficiency for the siRNA was elevated in a dose- and time- dependent fashion. Relative expression of SDF-1 and its receptor, CXCR4 and CXCR7 mRNA to ・-actin was reduced in a dose-dependent manner and hypoxia in a real-time RT-PCR study. Ultra-structure of A6K was transformed to micelle formation when siRNA was mixed.Lipid-like self assembling peptide, A6K includes genes in the micelle due to hydrophilic tail. This transfection is a novel and stable method with lower cytotoxity. In conclusion, SDF-1 and its receptor-mediated cell signal pathway is enhanced in hypoxic condition.