| 张宇虹,夏稻子,礼广森,武俊,由悦,黄冬梅,薄华颖,胡滨,毛鑫,周红艳.超声靶向破坏微泡技术介导shRNA抑制小鼠肝癌细胞株JNK1基因表达[J].中国医学影像技术,2014,30(8):1131~1135 |
| 超声靶向破坏微泡技术介导shRNA抑制小鼠肝癌细胞株JNK1基因表达 |
| Inhibitory effects of shRNA on expression of JNK1 in mouse hepatocellular carcinoma cell lines mediated by ultrasound-targeted microbubble destruction |
| 投稿时间：2014-05-11 修订日期：2014-07-14 |
| DOI： |
| 中文关键词: 超声检查 微泡 肝肿瘤 转染 |
| 英文关键词:Ultrasonography Microbubble Liver neoplasms Transfection |
| 基金项目:国家自然科学基金（81250018、81371582）、辽宁省科技厅科技计划项目（2012225021）。 |
| 作者 | 单位 | E-mail | | 张宇虹 | 大连医科大学附属第二医院超声科, 辽宁 大连 116023 | zhangyh_66@163.com | | 夏稻子 | 大连医科大学附属第二医院超声科, 辽宁 大连 116023 | | | 礼广森 | 大连医科大学附属第二医院超声科, 辽宁 大连 116023 | | | 武俊 | 大连医科大学附属第二医院超声科, 辽宁 大连 116023 | | | 由悦 | 大连医科大学附属第二医院超声科, 辽宁 大连 116023 | | | 黄冬梅 | 大连医科大学附属第二医院超声科, 辽宁 大连 116023 | | | 薄华颖 | 大连医科大学附属第二医院超声科, 辽宁 大连 116023 | | | 胡滨 | 大连医科大学附属第二医院超声科, 辽宁 大连 116023 | | | 毛鑫 | 大连医科大学附属第二医院超声科, 辽宁 大连 116023 | | | 周红艳 | 大连医科大学附属第二医院超声科, 辽宁 大连 116023 | |
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| 中文摘要: |
| 目的 探讨应用超声靶向破坏微泡（UTMD）技术介导shRNA抑制小鼠肝癌细胞株JNK1基因表达的能力。方法 构建并筛选shRNA最佳表达载体。体外培养肝癌细胞Hca-F，共分为5组：A组为空白对照组；B组为shRNA质粒组；C组为脂质体组；D组为超声微泡+超声辐照组；E组为脂质体+超声微泡+超声辐照组。应用倒置荧光显微镜观察转染率；荧光定量PCR检测JNK1的mRNA水平；Western-Blot检测JNK1的蛋白质表达。结果 获得了shRNA干扰效果最好的表达载体。各组转染率比较：E组均大于B、C、D组（P均<0.05）；C、D组之间差异无统计学意义（P>0.05）。荧光定量PCR和Western-Blot检测各组JNK1mRNA和蛋白表达比较：E组的JNK1mRNA和蛋白表达水平均最低（P均<0.05）。结论 脂质体转染法与UTMD技术结合可以提高小鼠肝癌细胞株JNK1 shRNA的转染效率，并能够增强基因表达的抑制效果。 |
| 英文摘要: |
| Objective To evaluate the inhibitory effects of shRNA on expression of JNK1 in mouse hepatocellular carcinoma cell lines mediated by ultrasound microbubble (UTMD). Methods The best shRNA vector was built and screened. The hepatocellular cell lines were cultured in vitro and divided into five groups: Group A (control group), group B (shRNA plasmid group), group C (lipofection group), group D (ultrasound microbubbles combined with ultrasound irradiation group), group E (lipofection combined with ultrasound microbubbles and ultrasound irradiation group. The transfection rate was observed by inverted flurescence microscope. The expression levels of JNK1 mRNA and protein were evaluated by flurescence quantitative PCR and Western Blot respectively. Results The best shRNA vector was established. Transfection rate of group E was larger than that of group B, C and D (all P<0.05) and there was no significant difference between group C and D (P>0.05). Expression levels of JNK1 mRNA and protein were lowest in group E (all P<0.05). Conclusion The transfection rate of JNK1 shRNA can be improved through the combination of lipofection and UTMD in mouse hepatocellular cell lines, therefore enhencing the inhibitory effects of gene expression. |
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