Efficient GLP-1 gene delivery using two-step transcription amplification plasmid system with a secretion signal peptide and arginine-grafted bioreducible polymer.
Efficient GLP-1 gene delivery using two-step transcription amplification plasmid system with a secretion signal peptide and arginine-grafted bioreducible polymer.
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DOI：
10.1016/j.jconrel.2011.09.072
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发表时间：
2012-01-30
期刊：
影响因子：
--
通讯作者：
Kim SW
中科院分区：
文献类型：
--
作者：
Kim TI;Lee M;Kim SW
结合信号肽（SP）和两步转录扩增（TSTA）系统的优势，构建了由pβ-Gal4-p65和pUAS-SP-GLP-1组成的编码双质粒（pDNA）系统（TSTA (SP-GLP-1)），以促进表达GLP-1的产生和分泌。利用精氨酸接枝生物可还原聚合物（ABP）作为基因载体，研究了其作为GLP-1基因传递系统的潜力。它们的多聚体尺寸约为140纳米，zeta电位值为20毫伏。ABP没有显示与PEI25k相反的细胞毒性。RT-PCR实验发现，与单pDNA系统（pβ-GLP-1）相比，TSTA-SP pDNA系统的GLP-1基因转录水平升高。GLP-1 ELISA检测还发现，TSTA (SP-GLP-1) pDNA系统的GLP-1分泌量是pβ-GLP-1的2.7 ~ 3.4倍，是TSTA （GLP-1）的1.5 ~ 1.7倍。ABP基因载体系统GLP-1分泌水平较PEI25k增加2.5 ~ 3.5倍。将含有分泌GLP-1的转染培养基转移到NIT-1胰岛素瘤细胞中，ABP/TSTA （SP-GLP-1）复合培养基处理的细胞胰岛素分泌水平最高。因此，该系统可作为一种安全有效的GLP-1基因传递系统用于2型糖尿病的治疗。
Glucagon-like peptide (GLP-1) encoding dual plasmid (pDNA) system (TSTA (SP-GLP-1)) which is composed of pβ-Gal4-p65 and pUAS-SP-GLP-1 was constructed to improve the production and secretion of expressed GLP-1 by combining the advantages of signal peptide (SP) and two-step transcription amplification (TSTA) system. Its potential for GLP-1 gene delivery system was investigated with employment of arginine- grafted bioreducible polymer (ABP) as a gene carrier. Their polyplexes have about 140 nm-sizes and 20 mV Zeta-potential values. ABP showed no cytotoxicity contrary to PEI25k. It was found in RT-PCR experiments that TSTA-SP pDNA systems showed increased GLP-1 gene transcription level in comparison with mono pDNA system (pβ-GLP-1). It was also observed in GLP-1 ELISA that GLP-1 secretion level of TSTA (SP-GLP-1) pDNA system was 2.7–3.4 times higher than those of pβ-GLP-1 and 1.5–1.7 times than TSTA (GLP-1). Additionally, 2.5–3.5 folds increased level of GLP-1 secretion was found in ABP gene carrier system in comparison with PEI25k. When transfection medium containing secreted GLP-1 was transferred to NIT-1 insulinoma cells, the highest secretion level of insulin was induced in ABP/TSTA (SP-GLP-1) polyplex medium-treated cells. Therefore, this novel system could be utilized as a safe and efficient GLP-1 gene delivery system for type 2 diabetes therapy.
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影响因子：
4.7
作者：
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