全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...
-  2015 

超声联合微泡介导缺氧诱导因子-1α基因转染293T细胞的实验研究 Experimental Study of Hypoxia Inducible Factor-1 Aplha Transfected into 293T Cells by Ultrasound-Mediated Microbubble Destruction

Keywords: 超声,微泡,基因转染,HIF-1α

Full-Text   Cite this paper   Add to My Lib

Abstract:

目的:探讨超声联合微泡造影剂介导缺氧诱导因子(HIF)-1α基因转染人胚肾293T细胞的转染效率。方法:将6孔板中的293T细胞悬液分为3组,A组:质粒+超声辐照组,加入质粒,终浓度为15μg/孔;B组:载基因微泡+超声辐照组,加入载基因质粒微泡混合液,质粒的终浓度为15μg/孔,微泡的终浓度为200μl/孔;C组:对照组,每孔中仅加入单纯质粒DNA,终浓度15μg/孔。A组和B组均接受超声辐照,超声照射条件为连续波,声强1.5 W/cm2,辐照时间30s。超声辐照转染48h后,用荧光显微镜和流式细胞仪分别定性定量观察各组细胞基因的转染效率。结果:转染48h后,荧光显微镜观察到转染成功的细胞内发出绿色荧光,流式细胞仪检测A组、B组、C组的基因转染效率分别为(5.6±0.5)%、(30.5±1.0)%、(0.1±0.1)%。结论:超声辐照联合微泡能够介导治疗性基因的体外细胞转染,为冠心病心肌缺血的HIF-1α基因治疗奠定基础

References

[1]  Kowalczuk L,Boudinet M,El Sanharawi M,et al.In vivo gene transfer into the ocular ciliary muscle mediated by ultrasound and microbubbles[J].Ultrasound Med Biol,2011,37(11):1 814-1 827.
[2]  Giaccia A,Siim BG,Johnson RS.HIF-1as a target for drug development[J].Nat Rev Drug Discov,2003,2(10):803-811.
[3]  聂金,戴爱国,胡瑞成.缺氧诱导因子脯氨酸羟化酶的调控与缺血缺氧性疾病[J].国际病理科学与临床杂志,2007,27(1):53-56.
[4]  Tsunoda S,Mazda O,Oda Y,et al.Sonoporation using microbubble BR14 promotes pDNA/siRNA transduction to murine heart[J].Biochem Biophys Res Commun,2005,336(1):118-127.
[5]  Bai WK,Wu ZH,Shen E,et al.The improvement of liposome-mediated transfection of pEGFP DNA into human prostate cancer cells by combining low-frequency and low-energy ultrasound with microbubbles[J].Oncol Rep,2012,27(2):475-480.
[6]  王璋,司良毅.低氧诱导因子1与心血管疾病[J].心脏杂志,2004,16(3):284-286.
[7]  Zhang Q,Wang Z,Ran H,et al.Enhanced gene delivery into skeletal muscles with ultrasound and microbubble techniques[J].Acad Radiol,2006,13(3):363-367.

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133