微小RNA传递载体聚乙二醇-b-聚赖氨酸的合成及细胞毒性研究

任海峰; 赵英魁; 杨峰; 俞媛; 马志强

doi:10.3969/j.issn.1006-0111.2014.05.013

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微小RNA传递载体聚乙二醇-b-聚赖氨酸的合成及细胞毒性研究

任海峰, 赵英魁, 杨峰, 俞媛, 马志强

文章导航 > 药学实践与服务 > 2014 > 32(5): 362-365,373

任海峰, 赵英魁, 杨峰, 俞媛, 马志强. 微小RNA传递载体聚乙二醇-b-聚赖氨酸的合成及细胞毒性研究[J]. 药学实践与服务, 2014, 32(5): 362-365,373. doi: 10.3969/j.issn.1006-0111.2014.05.013

引用本文:

REN Haifeng, ZHAO Yingkui, YANG Feng, YU Yuan, MA Zhiqiang. Study on synthesis and cyto-toxicity of polyethylene glycol-b-polylysine as potencial miRNA carrier[J]. Journal of Pharmaceutical Practice and Service, 2014, 32(5): 362-365,373. doi: 10.3969/j.issn.1006-0111.2014.05.013

Citation:

REN Haifeng, ZHAO Yingkui, YANG Feng, YU Yuan, MA Zhiqiang. Study on synthesis and cyto-toxicity of polyethylene glycol-b-polylysine as potencial miRNA carrier[J]. Journal of Pharmaceutical Practice and Service, 2014, 32(5): 362-365,373. doi: 10.3969/j.issn.1006-0111.2014.05.013

微小RNA传递载体聚乙二醇-b-聚赖氨酸的合成及细胞毒性研究

doi: 10.3969/j.issn.1006-0111.2014.05.013

第二军医大学药学院, 上海 200433

Study on synthesis and cyto-toxicity of polyethylene glycol-b-polylysine as potencial miRNA carrier

School of Pharmacy, Second Military Medical University, Shanghai 200433, China

摘要: 目的制备微小RNA(miRNA)传递载体聚乙二醇-b-聚赖氨酸(PEG-b-PLL),并且选用模型细胞对其进行毒性考察。方法通过核磁谱测定聚赖氨酸的聚合度,用4%琼脂糖凝胶电泳检测聚复合物对miRNA的包封,用动态光散射法测定制备所得聚复合物粒径、多分散性指数(PDI),Zeta电位,用荧光标记的FAM-hsa-miR-15a进行包封率的测定,采用CKK-8试剂盒测定K562细胞(人慢性淋巴白血病细胞)活力考察PEG-b-PLL的细胞毒性。结果用PEG-b-PLL制备的聚阳离子胶束/miRNA复合物特性符合应用要求,并且细胞毒性显著低于聚乙烯亚胺(PEI),lipo2000。结论离子聚合物载体PEG-b-PLL符合应用要求,同时毒性较低,有望成为基因转染的有效载体。
- 非病毒载体 /
- 聚赖氨酸 /
- 微RNA(miRNA) /
- 细胞毒性
Abstract: Objective To synthesize miRNA carrier PEG-b-PLL and to testify the stability, encapsulation efficiency and cytotoxicity of its complexes. Methods H¹NMR was used to determine the degree of polymerization of PLL, 4% agarose gel electrophoresis was used to determine entrapment of the polyer to the miRNA; then dynamic light scattering(DLS) was used to measure the hydrodynamic parameter such as size, polydispersity index(PDI) and zeta potential of the polyplexes. The entrapment efficiency was determined by ultraviolet-visible spectrophotometer, and finally, the cyto-toxicity of PEG-b-PLL was evaluated by CKK-8 kit with K562 cell lines. Results The characteristics indicated polyplexes prepared by PEG-b-PLL and miRNA fulfill the demand of being the gene carrier of miRNA because of low cyto-toxicity, high encapsulation efficiency and stability. Conclusion The miRNA carrier PEG-b-PLL had good character and low cyto-toxicity. It showed considerable potential as an efficient miRNA carrier.
- non-viral vector /
- polyethylenimine /
- miRNA /
- cyto-toxicity

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微小RNA传递载体聚乙二醇-b-聚赖氨酸的合成及细胞毒性研究

doi: 10.3969/j.issn.1006-0111.2014.05.013

第二军医大学药学院, 上海 200433

收稿日期: 2014-02-21
修回日期: 2014-06-28

关键词:

摘要: 目的制备微小RNA(miRNA)传递载体聚乙二醇-b-聚赖氨酸(PEG-b-PLL),并且选用模型细胞对其进行毒性考察。方法通过核磁谱测定聚赖氨酸的聚合度,用4%琼脂糖凝胶电泳检测聚复合物对miRNA的包封,用动态光散射法测定制备所得聚复合物粒径、多分散性指数(PDI),Zeta电位,用荧光标记的FAM-hsa-miR-15a进行包封率的测定,采用CKK-8试剂盒测定K562细胞(人慢性淋巴白血病细胞)活力考察PEG-b-PLL的细胞毒性。结果用PEG-b-PLL制备的聚阳离子胶束/miRNA复合物特性符合应用要求,并且细胞毒性显著低于聚乙烯亚胺(PEI),lipo2000。结论离子聚合物载体PEG-b-PLL符合应用要求,同时毒性较低,有望成为基因转染的有效载体。