脂质体载体转染GFP质粒在b16黑色素瘤细胞和黑色素瘤模型中的应用

Lapkina E.Z., Esimbekova A.R., Palkinа N.V.
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引用次数: 0

摘要

摘要背景:由于细胞内脂质膜和扩散屏障的屏障作用,质粒DNA通过被动过程渗透到细胞核是无效的。一个有效的基因传递系统应该确保与靶细胞结合,通过内吞作用穿透,防止降解,并最终将表达质粒传递到细胞核。转染后药物的分布途径对药物靶向作用的实现起着至关重要的作用。在我们的实验中,使用报告蛋白GFP的质粒DNA来研究其分布途径。目的:评价GFP报告蛋白在C57Bl6小鼠B16黑色素瘤细胞和远端器官中的分布,作为确定转染基因工程结构在体内肿瘤诊断和治疗中的有效性的方法。材料与方法:对皮肤黑色素瘤细胞系B16进行体外研究。用报告蛋白GFP质粒转染细胞。在体内,本研究以C57Bl/6系成熟雌性小鼠为实验对象,将肿瘤细胞移植到雌性小鼠体内,采用Lipofectamine 3.0和Invivofectamine两种商业转染剂腹腔注射GFP质粒。使用细胞显像站fluid评估转染器官和肿瘤细胞的效率。结果:B16黑色素瘤细胞存活率高,48h内可获得GFP+ B16黑色素瘤细胞。我们发现,通过腹腔注射含有GFP质粒的脂复合物,无论是使用Lipofectamine 3.0转染剂还是Invivofectamine,都能有效地融入肝脏和肾脏实质细胞。GFP报告蛋白在肝脏、肾脏和肿瘤结细胞中的渗透和滞留。结论:GFP质粒转染C57Bl6小鼠肝、肾、黑色素瘤B16肿瘤结细胞的有效性已得到证实。所获得的数据表明,利用转染剂渗透到基因工程结构的靶细胞中,用于体内肿瘤的诊断和治疗具有广阔的前景。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
TRANSFECTION OF GFP PLASMIDS BY LIPOSOMAL CARRIERS IN B16 MELANOMA CELLS AND MELANOMA MODELS IN VIVO
Abstract. Background: Penetration of plasmid DNA into the nucleus by a passive process is ineffective due to the barrier effect of lipid membranes and diffusion barriers inside the cell. An effective gene delivery system should ensure binding to the target cell, penetration by endocytosis, protection from degradation and, ultimately, delivery of the expression plasmid to the nucleus. The distribution routes of transfected agents play a crucial role in achieving the targeted effect of drugs. In our experiment, plasmid DNA of the reporter protein GFP was used to study the distribution pathways. Aim: to evaluate the distribution of GFP reporter protein in B16 melanoma cells and distant organs of C57Bl6 mice as a method for determining the effectiveness of transfection of genetically engineered structures for the diagnosis and therapy of tumors in vivo. Materials and methods: An in vitro study was conducted on skin melanoma cell lines B16. Cells were transfected with a plasmid of the reporter protein GFP. In vivo, the study was conducted on mature female mice of the C57Bl/6 line, to which tumor cells were transplanted and GFP plasmid was intraperitoneally injected using commercial transfectants Lipofectamine 3.0 and Invivofectamine. The efficiency of transfection in organ and tumor cells was evaluated using the cell imaging station Floid. Results: A high survival rate of B16 melanoma cells was established and GFP+ B16 melanoma cells were obtained within 48 hours. We found that with intraperitoneal administration of a lipocomplex containing GFP plasmid, both with the use of Lipofectamine 3.0 transfectant and Invivofectamine, effective incorporation into liver and kidney parenchyma cells occurs. The penetration and retention of GFP reporter protein in liver, kidney and tumor node cells was revealed. Conclusion: The effectiveness of transfection of GFP plasmid into liver, kidney, and melanoma B16 tumor node cells of C57Bl6 mice has been proven. The data obtained indicate the prospects for the use of transfecting agents for penetration into target cells of genetically engineered structures for the purpose of diagnosis and therapy of tumors in vivo.
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