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/hTERT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pEGFP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。
Abstract
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 1H-NMR was used to characterize 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 evaluated 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 cytotoxicity 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 transfection 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 polycationic carrier as shown by in vitro evaluation, has the potential to become a novel gene vector in the therapy of prostate cancer.
关键词
聚阳离子载体 /
前列腺癌 /
miR-15-a
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Key words
polycationic carrier /
prostate cancer /
miR-15-a
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中图分类号：
R944
R969
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参考文献
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脚注
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基金
国家自然科学基金资助项目(81201809);浙江省公益性技术应用研究计划资助项目(2015C37125);浙江省自然科学基金资助项目(LQ12H30005);嘉兴市科技计划资助项目(2012AY1075-3, 2015AY23064);“十二五”浙江省高校重点学科(药理学)资助项目
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