| 汪朝霞,王志刚,朱叶锋,李巧,康娟.超声声学造影剂介导肝细胞生长因子基因转染的体外实验[J].中国医学影像技术,2009,25(6):935~937 |
| 超声声学造影剂介导肝细胞生长因子基因转染的体外实验 |
| Contrast agents enhanced HGF gene transfection in vitro |
| 投稿时间：2008-12-02 修订日期：2009-02-13 |
| DOI： |
| 中文关键词: 造影剂 基因转染 超声检查 |
| 英文关键词:Contrast media Gene transfection Ultrasonography |
| 基金项目:863计划(2006AA02Z4FO),国家自然科学基金重点项目(30430230),重庆市自然科学基金重点项目(CSCT,2005BA5024)。 |
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| 中文摘要: |
| 目的 研究超声声学造影剂促进肝细胞生长因子(HGF)基因在大鼠肝星状细胞株(HSC-T6)中的转染效果。方法 将HSC-T6接种在24孔板中,随机分为5组:①空白对照组(C);②单纯质粒组(P);③载基因超声声学造影剂组(M);④质粒＋超声组(P+U);⑤载基因超声声学造影剂＋超声组(M+U)。荧光显微镜及流式细胞仪检测pIRES2-EGFP/HGF在细胞内的表达及转染效率;MTT法检测该方法对HSC-T6生长活性的影响;Western Blotting检测HGF蛋白的表达。结果 M+U组的绿色荧光强度及转染率均高于其他各组,并对细胞活性无明显影响,且HGF蛋白的表达均高于其他各组(P＜0.05)。结论 超声声学造影剂可提高基因在体外培养HSC-T6的转染效率,并对细胞活性无明显影响,为提高非病毒载体的转染效率提供一个新策略。 |
| 英文摘要: |
| Objective To study the transfection effects of contrast agents enhanced HGF gene in hepatic stellate cell (HSC-T6). Methods HSC-T6 was seeded in the 24-well plates, and divided into 5 groups randomly. The first group was regarded as contrast group, and the second group was given proper dose of plasmid (pIRES2-EGFP/HGF). Loaded-gene contrast agents were added in the third group, while pulsed-ultrasound exposed liposome was added in the fourth group. Loaded-gene contrast agents were added in the fifth group that exposed to pulsed-ultrasound, too. Expression of the report gene EGFP was observed on fluorescent microscopy and quantified with FACS analysis. The viability of HUVEC was measured with MTT assay. Western Blotting was used to detect the expression of HGF peptide. Results Green fluorescence intensity and transfection efficiency in the fifth group were higher than that in other groups, and no significant difference of cell viability was found. HGF protein in the fifth group was more than that in other groups. Conclusion Ultrasound microbubbles can enhance the efficiency of gene transfection without obvious damage to cell viability in HSC-T6. This method offers a new strategy to increase the transfection efficiency of non-viral genetic vector. |
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