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超声微泡介导EGFP质粒转染视网膜母细胞瘤细胞与传统转染方法效率对比
Experimental Research of Transfection Efficiency for EGFP Plasmid Transfected into Retinoblastoma Cells by Ultrasound Microbubble Intensifier
【摘要】 目的探讨超声微泡造影剂在一定能量的超声波辐照下,介导EGFP质粒转染视网膜母细胞瘤(RB)细胞的效率及可行性方法将RB细胞分为6组,1组以一定能量的超声波辐照,2组加适当剂量的微泡造影剂,3组加入质粒,4组加入质粒与微泡,5组加入质粒、微泡,并用一定能量的超声辐照,6组予脂质体与质粒。转染24~48h后观察EGFP表达,并用RT-PCR进行检测。同时对1、2组予以染色。结果超声微泡介导的DNA质粒对RB细胞的转染效率,与脂质体介导的质粒转染效率相似,明显高于其他实验组。一定能量和时间的超声波辐照,及适当浓度的微泡,对RB细胞的活性无明显抑制。结论利用低频率和一定能量的超声击碎携带EGFP质粒的超声微泡造影剂,能够有效地提高DNA质粒在RB细胞中的转染效率。
【Abstract】 Objective To investigate whether ultrasound-mediated microbubble destruction could effectively deliver plasmid to retinoblastoma cells.Methods The cultured retinoblastoma cells were divided into six groups.The first group was exposed to ultrasound and the second group was given proper dose of microbubbles.We added this plasmid to the third group and plasmid with microbubbles to the forth group.Plasmid,microbubbles and ultrasound were applied to the fifth group and plasmid with liposome to the sixth group.After 24 to 48 hours,the EGFP expression in the retinoblastoma(RB) cells were detected by fluorescence microscopy and RT-PCR.Trypanblue staining were adopted to measure the cell vitality of the first and second group.Results The infection efficiency of the fifth group were similar to that of the sixth group,and were both higher than other groups.Ultrasound used in the experiment did not harm to the cells neither did the microbubbles.Conclusions The EGFP expression in RB cells were increased with the administration of ultrasound-mediated microbubbles destruction.
- 【文献出处】 中国超声医学杂志 ,Chinese Journal of Ultrasound in Medicine , 编辑部邮箱 ,2006年08期
- 【分类号】R739.7
- 【被引频次】26
- 【下载频次】195