摘要
目的:构建含人SOX(SRY related high mobility group box)18的重组表达质粒,并研究其对血管内皮细胞增殖能力的影响。方法:通过RT-PCR技术从血管内皮细胞中扩增SOX18编码基因序列,将其插入真核表达载体pSec Tag-2B构建重组表达质粒pST-SOX18,分别转染人胚肾上皮细胞HEK293T和血管内皮细胞,通过免疫印迹技术检测SOX18的蛋白表达情况;进一步通过CCK-8实验检测SOX18对血管内皮细胞增殖能力的影响。结果:成功构建重组质粒p ST-SOX18,重组蛋白能够在HEK293T和血管内皮细胞中表达;SOX18的过表达促使血管内皮细胞合成血管内皮生长因子(VEGF),促进血管内皮细胞增殖。结论:SOX18重组蛋白能够促进血管内皮细胞增殖。
Objective:To construct the recombinant plasmid containing SRY related high mobility group box 18(SOX18)gene and investigate the effect of the recombinant protein on angiogenesis of vascular endothelial cells. Methods:The whole sequence of SOX18 gene that amplified from human umbilical vein endothelial cells(HUVEC)by reverse transcription-polymerase chain reaction(RTPCR)were inserted into the expression plasmid p Sec Tag-2B to construct the recombinant vector,and termed as pST-SOX18. pSTSOX18 was then transfected into the HEK293 T and vascular endothelial cells,respectively. Subsequently,Western blotting was carried out to determine the expression of the recombinant protein. CCK-8 assay was used to assess the effect of pST-SOX18 on proliferation of vascular endothelial cells. Results:The recombinant plasmid pST-SOX18 was constructed successfully. pST-SOX18 was successfully transfected into HEK293 T and vascular endothelial cells,target protein was determined by immunoblotting. pST-SOX18 transfection significantly promoted vascular endothelial cell proliferation in vitro. Conclusion:The recombinant plasmid expressing functional SOX18 was constructed successfully. SOX18 have a potent potential in promoting the proliferation of vascular endothelial cells.
出处
《南京医科大学学报(自然科学版)》
CAS
CSCD
北大核心
2018年第2期166-170,共5页
Journal of Nanjing Medical University(Natural Sciences)
基金
国家自然科学基金项目(81503368)
江苏省中医院青年项目(Y14011)
关键词
SOX18
载体构建
细胞增殖
SOX 18
plasmid construction
cell proliferation