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目的:探讨超声介导微泡破裂法促进外源基因的安全转移的方法,为临床上肿瘤等疾病的基因治疗研究提供新的思路。方法:以绿色荧光蛋白基因为标记基因,以人大肠癌细胞株SW480为靶细胞,分别以自制白蛋白微泡和声诺维(SonoVue)微泡作为载体,通过超声介导微泡破裂法促进绿色荧光蛋白GFP基因在人大肠癌细胞株的定向转染,以激光共聚焦显微镜来定性和半定量观察GFP在靶细胞的表达情况,以椎虫蓝染色法检测超声作用于细胞的安全性。结果:当超声的强度为0.75W/cm2,作用时间为40s时,对细胞比较安全;自制白蛋白微泡和声诺维微泡的浓度为10%时,分别达到最佳的基因转染效率,两者无显著性差异,但后者的相对表达强度较高。结论:超声介导微泡破裂法促进外源基因的转移是一种比较安全而有效的基因转染方法。
OBJECTIVE: To investigate the method of ultrasound-mediated microbubble rupture to promote the safe transfer of exogenous genes and to provide new ideas for gene therapy research on clinical diseases such as tumors. Methods: The green fluorescent protein (EGFP) gene was used as a marker gene. Human colorectal cancer cell line SW480 was used as target cell. The self-made albumin microbubbles and SonoVue microbubbles were used as vectors. Green fluorescent protein GFP gene was transfected into human colorectal cancer cell lines with confocal laser scanning microscopy to qualitatively and semi-quantitatively observe the expression of GFP in target cells. The safety of ultrasound on cells was detected by the method of vertebrate-blue staining. Results: When the intensity of ultrasound was 0.75 W / cm2 and the action time was 40 s, it was safer for the cells. When the concentration of homemade albumin microbubbles and sonicate microbubbles was 10%, the optimal gene transfection efficiency , No significant difference between the two, but the relative intensity of the latter higher. CONCLUSIONS: Ultrasound-mediated microbubble disruption promotes the transfer of exogenous genes is a safe and effective method of gene transfection.