| 王玉强 王飞 曹青 苏靖 盛净 陈书艳.PEI-Et输送特异性shRNA对大鼠肝细胞AGT 基因表达的影响[J].,2014,14(7):1267-1270 |
| PEI-Et输送特异性shRNA对大鼠肝细胞AGT 基因表达的影响 |
| Effects of Delivery of Specific shRNA using PEI-Et on AGT GeneExpression in Rat Liver Cells |
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| DOI： |
| 中文关键词: AGT RNA 干扰 BRL-3A 转染效率 细胞毒性 |
| 英文关键词: AGT RNAinterference BRL-3A Transfection efficiency Cytotoxicity |
| 基金项目:国家自然科学基金项目（81001416；81270205） |
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| 中文摘要: |
| 目的：研究交联小分子量聚乙烯亚胺衍生物PEI-Et对大鼠肝细胞（BRL-3A）的细胞毒性、转染效率和携带高血压相关基因血
管紧张素原（AGT）短发卡RNA(shRNA)沉默AGT 表达的能力。方法：MTT 法检测PEI-Et/shRNA 复合物对BRL-3A 细胞的毒
性，流式细胞术检测PEI-Et/shRNA复合物对BRL-3A细胞的转染效率，RT-PCR 和Western blot 检测PEI-Et/shRNA 对AGT 的基
因沉默效果。结果：在相同质量比(w/w)时PEI-Et/shRNA的细胞毒性小于PEI 25kDa/shRNA(P<0.01)，PEI-Et/shRNA在w/w 为30
时达到最高转染效率，高于PEI 25 kDa (P<0.01), PEI-Et/shRNA能高效沉默BRL-3A 细胞中AGT 基因的表达。结论：PEI-Et 在
BRL-3A 细胞中是一种低细胞毒性、高转染效率的非病毒基因载体( 与商业化的PEI 25kDa 比较)，能携带AGT shRNA 高效沉默
BRL-3A 细胞中AGT基因的表达，通过用PEI-Et/AGT shRNA来抑制AGT 的表达将为高血压的基因治疗提供一种新的思路。 |
| 英文摘要: |
| Objective:To synthesize cross-linked small-molecular-weight polyethylenimine derivative PEI-Et and investigate its
cytotoxicity, transfection efficiency and ability to delivery hypertension related gene angiotensinogen （AGT）short hairpin RNA
(shRNA) to silence AGT expression.Methods: MTT assay was used to measure the cytotoxicity of PEI-Et/shRNA complexes. Flow
cytometry was performed to investigate transfection efficiency of PEI-Et/shRNA in BRL-3A cells. RT-PCR and Western blot were used
to detect the AGT gene silencing effect of PEI-Et/shRNA.Results:PEI-Et/shRNA showed lower cytotoxicity than PEI 25kDa/shRNA at
the same weight ratio (w/w). Transfection results indicated that PEI-Et/shRNA displayed the highest transfection efficiency at w/w 30,
which was higher than PEI 25kDa/shRNA (P<0.01). PEI-Et/shRNA could efficiently inhibit the expression of AGT in BRL-3A cells.Conclusion:PEI-Et was a non-viral vector with much lower cytotoxicity and enhanced transfection efficiency than PEI 25kDa in BRL-3A
cells, and it could delivery AGT shRNA to efficiently silence AGT expression in BRL-3A cells. Therefore, PEI-Et/AGTshRNA would be
a promising tool for delivering AGT shRNA to BRL-3A cells for hypertension therapy. |
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