载miR-15-a基因纳米复合物的制备及其抗前列腺癌作用的体外研究

Construction of miR-15-a-Loaded Nano-Complex and Evaluation of Its Anti-Prostate Cancer Effect in Vitro

目的基于二硫键交联合成精氨酸-组氨酸(HRss)阳离子聚合物,构建新型纳米复合物HRss/miR-15-a,用于前列腺癌研究。方法利用~1H-NMR鉴定HRss2/miR-15-a载体的结构,通过电位粒度仪测定纳米复合物的电位和粒径,利用琼脂糖凝胶电泳考察载体对miR-15-a的包裹能力。以体外培养的前列腺癌干细胞RC-92a/h TERT为研究对象,CCK8法检测3种纳米复合物对细胞增殖的抑制作用,以p EGFP为报告基因考察基因转染效率,利用Transwell小室考察HRss/miR-15-a对细胞运动能力的影响。结果细胞毒性试验结果显示,载体材料对正常及肿瘤细胞毒性较低,可以包裹miR-15-a形成稳定的纳米复合物,HRss2/miR-15-a相较于HRss1/miR-15-a和HRss3/miR-15-a,转染效率更高,对前列腺癌干细胞的运动能力影响也更大。结论 HRss可以运载基因药物,体外评价效果最佳的HRss2,有潜力成为抗前列腺癌基因治疗中的新型载体。

OBJECTIVE To synthesize cationic polymers of arginine-histidine （HRss） based on disulfide crosslink and construct novel nano-complex HRss/miR-15-a, then study its anti-prostate cancer effect in vitro. METHODS I H-NMR was used to character- ize HRss2/miR-15-a. Zeta sizer was adopted to estimate the Zeta potential and particle size of the nano-complex. Gel electrophoresis was employed to determine the condensation capacity of HRssto miR-15-a. The cytotoxicity and transfection efficiency of HRss was e- valuated using prostate cancer stem-like cells （RC-92a/hTERT）. Transwell chambers were used to evaluate the influence of HRss/ miR-15-a against the motility of RC-92a/hTERT. RESULTS The results of cytotoxieity tests indicated that the carrier HRss2 had low toxicity in both normal cells and cancer cells, and the miR-15-a could be loaded in HRss2 to form stable nano-complex. The transfec- tion efficiency and inhibited motility of HRss2/miR-15-a against RC-92a/hTERT were statistically higher than those of HRss1/miR-15- a and HRss3/miR-15-a. CONCLUSION HRss may be useful for gene delivery, and HRss2, as the optimum polycationie carrier as shown by in vitro evaluation, has the potential to become a novel gene vector in the therapy of prostate cancer.