Luo J, Zhou X, Diao L, Wang Z
Department of Ophthalmology, The Second Affiliated Hospital of Chongqing University of Medical Sciences, Chongqing, China.
J Int Med Res. 2010 May-Jun;38(3):1005-15. doi: 10.1177/147323001003800327.
The transfection efficiency of wild-type p53 (wtp53) was investigated in retinoblastoma (RB) Y79 cells using an ultrasound microbubble technique. A human RB nude mouse xenograft tumour model was also used to investigate whether this technique could deliver wtp53 into solid tumours. Reverse transcription-polymerase chain reaction (RT-PCR) demonstrated that wtp53 was successfully transfected into Y79 cells in the plasmid with microbubbles and ultrasound group and in the plasmid with liposomes group, but not in the plasmid with ultrasound group or in the untreated control group. Flow cytometry showed that apoptosis was highest in the microbubbles and ultrasound group (25.58%) compared with the plasmid with liposomes group (19.50%), and the other two groups (< 10%). RT-PCR also showed that the wtp53 gene was successfully transfected into solid tumours in the plasmid with microbubbles and ultrasound group. This study provides preliminary evidence in support of a potential new approach to RB gene therapy.
利用超声微泡技术在视网膜母细胞瘤(RB)Y79细胞中研究野生型p53(wtp53)的转染效率。还使用人RB裸鼠异种移植瘤模型来研究该技术是否能将wtp53递送至实体瘤中。逆转录聚合酶链反应(RT-PCR)表明,在质粒与微泡及超声组和质粒与脂质体组中,wtp53成功转染至Y79细胞,但在质粒与超声组或未处理的对照组中未成功转染。流式细胞术显示,与质粒与脂质体组(19.50%)相比,微泡及超声组的凋亡率最高(25.58%),其他两组(<10%)。RT-PCR还表明,在质粒与微泡及超声组中,wtp53基因成功转染至实体瘤中。本研究为RB基因治疗的潜在新方法提供了初步证据。