Wnt5a involved in regulation of the biological behavior of hepatocellular carcinoma.
OBJECTIVE
Wnt5a has been shown to be involved in cancer progression in a variety of tumor types. Previous experimental studies have indicated that it has been shown to be down-regulated in hepatocellular carcinoma (HCC). The goal of this study was to explore the effect of Wnt5a overexpression in an HCC cell line.


METHODS
We transfected the human HCC cell line Huh7 with a pcDNA3.1-Wnt5a overexpression vector or an empty vector control. The integration of the plasmid DNA and the expression of Wnt5a in Huh7 cells were confirmed by real-time RT-PCR and Western blot. A plate colony formation test was used to calculate the clone formation rate and the cell cycle was analyzed by flow cytometry. The effect of Wnt5a overexpression on cell migration was studied in vitro using a scratch assay and in vivo by xenograft studies in nude mice.


RESULTS
Our results showed that in Huh7 cells with overexpression of Wnt5a, the fraction of cells in the G1 and S phases of the cell cycle was significantly increased compared with untransfected cells. In agreement with this finding, overexpression of Wnt5a was associated with a lower colony formation rate compared with control cells. In our xenograft studies, nude mice injected with Huh7 cells with overexpression of Wnt5a had decreased tumor volumes compared with controls. The vitro scratch assay revealed that Wnt5a overexpression cells had a diminished capacity for cell migration. Furthermore, we studied the expression of important proteins associated with Wnt5a signaling pathway, and it was found that Ror2 and E-cadherin were both increased in Huh7 cells with overexpression of Wnt5a, whereas p53 expression was unaffected.


CONCLUSIONS
Overexpression of Wnt5a in Huh7 cells was associated with decrease of cell proliferation and migration. Wnt5a may act as a tumor-suppressor gene in HCC, which works through the non-canonical Wnt signaling pathway by binding to the Ror2 and E-cadherin receptor.
Department of Pathology, General Hospital of Jinan Military Command.
Liquan
L
Bi
Department of Pathology, General Hospital of Jinan Military Command.
Xiaohong
X
Liu
Department of Pathology, General Hospital of Jinan Military Command.
Cuicui
C
Wang
Department of Pathology, General Hospital of Jinan Military Command.
Yongcheng
Y
Cao
Department of Pathology, General Hospital of Jinan Military Command.
Ruiqi
R
Mao
Department of Pathology, General Hospital of Jinan Military Command.
Peifeng
P
Li
Department of Pathology, General Hospital of Jinan Military Command.
Ming
M
Geng
D000818:Animals; D015153:Blotting, Western; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000378:metabolism* / Q000473:pathology*; D045744:Cell Line, Tumor; D002465:Cell Movement; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000502:physiology; D014162:Transfection; D051153:Wnt Proteins / Q000235:genetics / Q000378:metabolism*; D000071818:Wnt-5a Protein
D011518:Proto-Oncogene Proteins; C085389:WNT5A protein, human; D051153:Wnt Proteins; D000071818:Wnt-5a Protein
Hepatocellular carcinoma; Huh7; Wnt5a; migration; proliferation