title: Inhibitory effects of shRNA on expression of JNK1 and migration and invasion in mouse hepatocellular carcinoma cell lines mediated by ultrasound-targeted microbubble destruction.
abstract: OBJECTIVE
  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).
  
  
  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<0.05). There was no significant difference between the group of Lipofectamine and the group of UTMD (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<0.05). CCK-8 showed that cell viability decreased most in the group of Lipofectamine and UTMD (all P<0.05); Transwell assay showed that the abilities of migration and invasion decreased most in the group of Lipofectamine and UTMD (all P<0.05).
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Dignostic Ultrasound, Second Affiliated Hospital of Dalian Medical University, 116023 Dalian, PR China. Electronic address: zhangyh_66@163.com.
authors.forename: Yuhong
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Dignostic Ultrasound, Second Affiliated Hospital of Dalian Medical University, 116023 Dalian, PR China.
authors.forename: Jun
authors.initials: J
authors.lastname: Wu
authors.affiliation: Department of Dignostic Ultrasound, Second Affiliated Hospital of Dalian Medical University, 116023 Dalian, PR China.
authors.forename: Zihang
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Department of Dignostic Ultrasound, Second Affiliated Hospital of Dalian Medical University, 116023 Dalian, PR China.
authors.forename: Daozi
authors.initials: D
authors.lastname: Xia
authors.affiliation: Department of Dignostic Ultrasound, Second Affiliated Hospital of Dalian Medical University, 116023 Dalian, PR China.
authors.forename: Guangsen
authors.initials: G
authors.lastname: Li
authors.affiliation: Department of Dignostic Ultrasound, Second Affiliated Hospital of Dalian Medical University, 116023 Dalian, PR China.
authors.forename: Yue
authors.initials: Y
authors.lastname: You
authors.affiliation: Department of Dignostic Ultrasound, Second Affiliated Hospital of Dalian Medical University, 116023 Dalian, PR China.
authors.forename: Dongmei
authors.initials: D
authors.lastname: Huang
authors.affiliation: Department of Dignostic Ultrasound, Second Affiliated Hospital of Dalian Medical University, 116023 Dalian, PR China.
authors.forename: Huaying
authors.initials: H
authors.lastname: Bo
authors.affiliation: Department of Dignostic Ultrasound, Second Affiliated Hospital of Dalian Medical University, 116023 Dalian, PR China.
authors.forename: Bin
authors.initials: B
authors.lastname: Hu
authors.affiliation: Department of Pathology, Dalian Medical University, 116044 Dalian, PR China.
authors.forename: Jianwu
authors.initials: J
authors.lastname: Tang
mesh_terms: D000818:Animals; D015153:Blotting, Western; D006528:Carcinoma, Hepatocellular / Q000201:enzymology* / Q000235:genetics / Q000473:pathology*; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D015972:Gene Expression Regulation, Neoplastic; D008113:Liver Neoplasms / Q000235:genetics / Q000473:pathology; D008297:Male; D051379:Mice; D045423:Microbubbles; D048055:Mitogen-Activated Protein Kinase 8 / Q000378:metabolism*; D009361:Neoplasm Invasiveness; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism*; D014162:Transfection; D014465:Ultrasonics
chemical_list: D012333:RNA, Messenger; D034741:RNA, Small Interfering; D048055:Mitogen-Activated Protein Kinase 8
keywords: Hepatocellular carcinoma; JNK1; Lymphatic metastasis; Transfection; Ultrasound microbubble; shRNA