[Effects of ARNT2 gene on invasion and migration of human hepatocellular carcinoma HCCLM6 cell line].
OBJECTIVE
To investigate the effects of ARNT2 on invasion and migration of HCCLM6 cells.


METHODS
Four short hairpin oligos targeting to ARNT2 were s cloned into the pLVTHM vector. Lentiviral vectors shRNA-ARNT2i, pCMV-dR8.74 and pMD2G were cotransfected into 293T cells using Lipofectamine 2000. HCCLM6 was infected with virus supernatant. ARNT2 mRNA and protein expressions were detected using quantitative Real time-PCR and Western blot, respectively. The invasion and migration of HCCLM6 cells were evaluated using wound healing assay and cell invasion assay in vitro. Statistical analysis was performed with SPSS 16.0.


RESULTS
The relative mRNA levels of ARNT2 were 0.154+/-0.024, 0.860+/-0.145, 1.004+/-0.009 in shRNA-ARNT2i virus infected HCCLM6 cells, mock-infected cells and control vector virus infected cells (F = 113.14, P more than 0.01). The expression of ARNT2 at protein level was 16.45+/-1.6, 44.56+/-2.07 in the HCCLM6 cells infected with shRNA-ARNT2i virus and negative control vector virus, respectively (t = 18.58, P less than 0.01). The scrape wound of HCCLM6 cells infected with shRNA-ARNT2i virus healed faster than cells infected with control vector virus or mock-infected cells. The number of cells invading through Matrigel was higher in the HCCLM6 cells infected with shRNA-ARNT2i virus (13.25+/-1.04) than that in mock-infected HCCLM6 cells and the HCCLM6 cells infected with negative control vector virus (6.50+/-2.56, 6.75+/-2.05) (F = 29.645, P less than 0.01).


CONCLUSIONS
Inhibition of ARNT2 gene promotes the invasion and migration of HCCLM6 cells.
Li
Wei-Wei
WW
Liver Cancer Institute and Zhongshan Hospital, Fudan Unviersity, Shanghai 200032, China.
Wu
Wei-Zhong
WZ
Liang
Ying
Y
Xiao
Chuan-Li
CL
Tao
Zhong-Hua
ZH
Wang
Lu
L
Sun
Hui-Chuan
HC
Fan
Jia
J
D051784:Aryl Hydrocarbon Receptor Nuclear Translocator / Q000235:genetics* / Q000378:metabolism; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics* / Q000378:metabolism; D015153:Blotting, Western; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D045744:Cell Line, Tumor; D002465:Cell Movement; D015972:Gene Expression Regulation, Neoplastic; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D008113:Liver Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D009361:Neoplasm Invasiveness; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D014162:Transfection
D051784:Aryl Hydrocarbon Receptor Nuclear Translocator; D051792:Basic Helix-Loop-Helix Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; C490989:ARNT2 protein, human