Journal of Jilin University Medicine Edition ›› 2015, Vol. 41 ›› Issue (01): 21-26.doi: 10.13481/j.1671-587x.20150105

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Inhibition of expression of JNK1 in mouse hepatocellular carcinoma cell lines and cell migration and invasion mediated by ultrasound-targeted microbubble destruction

WANG Zihang1,2, ZHANG Yuhong2, XIA Daozi2, LI Guangsen2, WU Jun2, YOU Yue2, HUANG Dongmei2, BO Huaying2, HU Bin2, MAO Xin2   

  1. 1. Graduate School, Dalian Medical University, Dalian 116044, China;
    2. Department of Dignostic Ultrasound, Second Affiliated Hospital, Dalian Medical University, Dalian 116023, China
  • Received:2014-07-30 Published:2015-01-30

Abstract:

Objective To explore the inhibition of the expression of JNK1 in mouse hepatocellular carcinoma cell lines and cell migration and invasion mediated by ultrasound-targeted microbubble destruction (UTMD), and to clarify its mechanism. Methods The best short hairpin RNA(shRNA) vector was established and screened.The hepatocellular cell line Hca-F were cultured in vitro and divided into normal Hca-F cells group, shRNA plasmid group, and lipofection group, ultrasound microbubbles combined with ultrasound irradiation group, lipofection combined with ultrasound microbubbles and ultrasound irradiation group.The transfection rates were observed by inverted flurescence microscope.The expression levels of JNK1 mRNA and protein were detected by fluorescence quantitative PCR and Western blotting method.The activities of the cells in varions groups were detected by CCK-8 method.The abilities of migration in vitro of the cells in various groups were detected by Transwell assay. Results The transfection rate of the cells in lipofection combined with ultrasound microbubbles and ultrasound irradiation group was higher than those in shRNA plasmid group, lipofection group and ultrasound microbubbles combined with ultrasound irradiation group (all P<0.05);there was no signifant difference between lipofection group and ultrasound microbubbles combined with ultrasound irradiation group (P>0.05).The expression levels of JNK1 mRNA and protein in lipofection combined with ultrasound microbubbles and ultrasound irradiation group were lower than those in other groups (all P<0.05). The cell activity and the average amount of the cells passed the membrane in lipofection combined with ultrasound microbubbles and ultrasound irradiation group were lower than those in other groups (all P<0.05). Conclusion Combination of lipofection and UTMD in mouse hepatocellular cell lines can improve the transfection rate of JNK1 shRNA, therefore enhance the inhibitory effects on gene expression, the cell activity, migration and invasion.

Key words: ultrasound, microbubble, liver neoplasms,experimental, c-Jun N-terminal kinase, short hairpin RNA, transfection, lympatic metastasis

CLC Number: 

  • R735.7