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Inhibitory effects of shRNA on expression of JNK1 and migration and invasion in mouse hepatocellular carcinoma cell lines mediated by ultrasound-targeted microbubble destruction

脂质体 转染 细胞培养 小发夹RNA 细胞 基因敲除 分子生物学 生物 免疫印迹 载体(分子生物学) 基因 生物化学 重组DNA 遗传学
作者
Yuhong Zhang,Jun Wu,Zihang Wang,Daozi Xia,Guangsen Li,Yue You,Dongmei Huang,Huaying Bo,Bing Hu,Jianwu Tang
出处
期刊:Biomedicine & Pharmacotherapy [Elsevier BV]
卷期号:78: 1-7 被引量:2
标识
DOI:10.1016/j.biopha.2015.12.026
摘要

Abstract Aim The inhibitory effects on expression of JNK1 in mouse hepatocellular carcinoma cell lines and cell migration and invasion were mediated by ultrasound-targeted microbubble destruction (UTMD). Materials and methods The best shRNA vector was built and screened. The hepatocellular carcinoma cell lines were cultured in vitro and divided into five groups: the group of normal Hca-F cells, the group of shRNA plasmid (already selected from the above procedure), the group of Lipofectamine, the group of UTMD (ultrasound microbubbles combined with ultrasound exposure) and the group of Lipofectamine and UTMD. The transfection rate was observed by inverted fluorescence microscope. The expression levels of JNK1 mRNA and protein were evaluated by fluorescence quantitative PCR and Western Blot respectively. The cell proliferation was detected by CCK-8. The ability of migration and invasion in vitro was detected by transwell assay. Results The best shRNA vector was established. The comparison of the transfection rate: The group of Lipofectamine and UTMD was larger than that of the groups of shRNA plasmid, Lipofectamine lipofection and UTMD (all P P  > 0.05). The comparison of the expression levels of JNK1 mRNA and protein: Both of the mRNA and protein expression levels were lowest in the group of Lipofectamine and UTMD (all P P P Conclusion The transfection rate of JNK1 shRNA can be improved through the combination of lipofection and UTMD in mouse hepatocellular carcinoma cell lines, therefore enhancing the inhibitory effects of gene expression. The inhibitory effects of cell proliferation, migration and invasion can also be enhanced.
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