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

在目前的研究中，我们正在开发一种新颖的转染方法，用表面活性来利用短干扰RNA （siRNA）来利用纳米管。A6 K的稳定性与六种丙氨酸溶液和羟基头、赖氨酸、常规性中央转染剂试剂、siFECTOR和Lipofectamine 2000年进行比较。人类大鼠骨髓瘤腺细胞系、AtT-20或GH 3的细胞毒性是根据MTS的试验进行的。用FITC-labelled siRNA靶向SDF-1受体mRNA分析了转染有效性，用荧光活性和显微镜进行了分析。A6 K的超结构被观察到电子显微镜，A6 K的非细胞毒性水平（IC 50， 320.8 ·g/ml）明显高于sifECTOR （（IC 50， 35.6 ·g/ml）和Lipofectamine 2000 （IC 50， 22.4 ·g/ml）。siRNA在剂量和时间依赖的时尚中被提升的转染效率。SDF-1及其受体、CXCR 4和CXCR 7 mRNA的相对表达--actin在实时RT-PCR研究中减少了剂量-依赖性标记和hypoxia。当siRNA混合时，A6 K的超结构被转化为微结构，如脂质体本身组装肽，A6 K包括微结构中的基因到羟基尾部。这种转染是一种新颖而稳定的低细胞毒性方法。在整合中，SDF-1及其受体介导的细胞信号通路在hypoxic condition中增强。

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.