Feasibility and effect of ultrasound microbubble-mediated wild-type p53 gene transfection of HeLa cells
Feasibility and effect of ultrasound microbubble-mediated wild-type p53 gene transfection of HeLa cells
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DOI：
10.3892/etm.2012.528
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发表时间：
2012-06-01
影响因子：
2.7
通讯作者：
Wang, Zhi-Gang
中科院分区：
文献类型：
--
作者：
Chen, Wen-Juan;Xiong, Zheng-Ai;Wang, Zhi-Gang
基因疗法在治疗疾病方面大有希望。基因治疗的关键问题是有效载体的选择。超声介导微泡造影技术（UMMT）已在除宫颈癌外的多种肿瘤中显示出良好的应用前景。在本研究中，根据MTT法的结果，我们初步选择了超声强度为0.5 W/cm（2），超声暴露时间为30秒，微泡浓度为10%作为Wtp 53质粒转染HeLa细胞的最佳实验条件。为了进一步研究超声联合微泡的转染效率，使用RT-PCR分析检测p53的mRNA水平。质粒+微泡组和超声组的转染效率明显高于其他各组。转染wtp 53基因后，流式细胞仪分析显示HeLa细胞的细胞周期被阻滞在G1期。本研究的结果表明，UMMT，一种新的基因传递系统，提高了wtp 53基因的转染效率。此外，通过引入wtp 53，HeLa细胞的生长被阻止。本研究为宫颈癌的基因治疗提供了新的方向。
Gene therapy holds great promise for the treatment of diseases. The key problem of gene therapy is the choice of an effective vector. Ultrasound-mediated microbubble technique (UMMT) has already shown promising applications in numerous types of tumors apart from cervical carcinoma. In the present study, according to the results of an MTT assay, we initially chose an ultrasound intensity of 0.5 W/cm(2), an ultrasound exposure time of 30 sec and a microbubble concentration of 10% as the optimum experimental condition for Wtp53 plasmid transfection into HeLa cells. To further investigate the transfection efficiency of ultrasound combined with microbubbles, RT-PCR analysis was used to examine the mRNA level of p53. The transfection efficiency in the plasmid plus microbubbles and ultrasound group was significantly higher than that of the other groups. Following transfection of the wtp53 gene, flow cytometric analysis showed that the cell cycle of HeLa cells was arrested in the G1 phase. The results of the present study suggest that UMMT, a new gene delivery system, increases the transfection efficiency of the wtp53 gene. Moreover, the growth of HeLa cells was arrested by introducing wtp53. This study may afford a new trend for the gene therapy of cervical carcinoma.