| 邓鑫,周希瑗,王志刚.超声微泡介导EGFP 质粒对视网膜母细胞瘤细胞转染效率的实验研究[J].中国医学影像技术,2006,22(2):163~166 |
| 超声微泡介导EGFP 质粒对视网膜母细胞瘤细胞转染效率的实验研究 |
| Experimental study of transfection eff iciency for EGFP plasmid transfected into retinoblastoma cells by ultrasound microbubble intensif ier |
| 投稿时间：2005-09-11 修订日期：2005-11-09 |
| DOI： |
| 中文关键词: 超声检查 造影剂,微泡 视网膜母细胞瘤 基因治疗 |
| 英文关键词:Ult rasonography Cont rast media , microbubbles Retinoblastoma Gene therapy |
| 基金项目:本研究受国家自然科学基金重点(30430230) 及面上(30370402) 项目资助。 |
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| 中文摘要: |
| 目的 探讨不同浓度的超声微泡造影剂,在不同声强的超声辐照下,介导DNA 质粒转染视网膜母细胞瘤(RB)细胞的效率及可行性,为实现外源基因高效、定向的转移奠定基础。方法 将培养的RB 细胞分别予以超声条件为0. 25 ,0. 5 , 0. 75 , 1. 0 , 1. 25 W/ cmp>2p> , 60 s 的连续波辐照,微泡造影剂浓度为1 % , 10 % , 20 % ,30 % ,以筛选出对RB 细胞活性无明显抑制的最适超声声强、辐照时间和微泡浓度。根据以上筛选条件,转染EGFP 基因入RB 细胞,24～48 h 后,在荧光显微镜下观察EGFP 表达情况,并用RT2PCR 对EGFP mRNA 进行半定量检测。结果 声强< 0. 75 W/ cm2 (60 s) ,以及微泡浓度< 20 %时,对RB 细胞的活性无明显抑制。当微泡浓度10 % ,超声声强为0. 5 W/ cmp>2p> 或0. 75 W/ cmp>2p> 时,介导的DNA质粒对RB 细胞转染具有较高的转染效率,明显高于其他实验组。超声声强为0. 5 W/ cmp>2p> 或0. 75 W/ cmp>2p> 介导的转染效率,在统计学上差异无显著性意义。结论 浓度适当的微泡在优化的声强条件下,能够有效地提高DNA 质粒在RB 细胞中的转染效率。 |
| 英文摘要: |
| Objective To investigate the efficiency and feasibility of DNA plasmid t ransfected into retinoblastoma ( RB)cells by microbubbles of different dosage and ultrasound with various intensity , and to afford a base for effective and direc-tional gene delivery. Methods In order to prevent the harm to RB cells , the optimum ultrasound intensity and microbubblesdosage were screened. The cultured retinoblastoma cells were exposed to different ultrasound with intensity of 0. 25 , 0. 5 ,0. 75 , 1. 0 and 1. 25 W/ cm2respectively , and the time were all 60 s. The other RB cells were exposed to microbubbles ofvarious dosage of 1 % , 10 % , 20 % and 30 % respectively. According to the optimum ultrasound intensity and microbubblesdosage , EGFP gene were t ransfected into RB cells. Af ter 24 to 48 hours , the EGFP expression in the RB cells were detectedby fluorescence microscopy and RT-PCR. Results Ult rasound with the intensity under 0. 75 W/ cm2 (60 s) , and microbub-bles with the dosage under 20 % , did no harm to RB cells. The t ransfected efficiency of the two groups (0. 5 W/ cm2 and0. 75 W/ cm2 ) were both higher than other group s. Conclusion The efficiency of DNA vector t ransfected into RB cells couldbe increased obviously by optimum ultrasound intensity and microbubbles dosage. |
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