Downregulation of microRNA-122 promotes proliferation, migration, and invasion of human hepatocellular carcinoma cells by activating epithelial-mesenchymal transition.

Onco Targets Ther 2016 6;9:2035-47. Epub 2016 Apr 6.

Department of Oncology, The Affiliated Jiangyin Hospital, School of Medicine, Southeast University, Jiangyin, Jiangsu, People's Republic of China.

Objective: To investigate the effects of microRNA-122 (miR-122) on proliferation, migration, and invasion in human hepatocellular carcinoma (HCC) cells by activating epithelial-mesenchymal transition (EMT) pathways.

Methods: miR-122 mimics, miR-122 inhibitors, relevant control oligonucleotides, and Wnt1 were transfected into HepG2 and huh7 cell lines which were then divided into six groups: miR-122 group, anti-miR-122 group, miR-negative control (NC) group, anti-miR-NC group, miR-122 + Wnt1 group, and miR-122 + vector group. The miR-122 expressions and mRNA expressions of Wnt1 and EMT-related genes (E-cadherin, vimentin, β-cadherin, and N-cadherin) were quantified by quantitative real-time polymerase chain reaction (qRT-PCR). Protein expression levels of Wnt1, E-cadherin, vimentin, β-cadherin, and N-cadherin were measured by Western blot. Cell proliferation, migration, and invasion were evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, wound-healing assay, and Transwell assay, respectively.

Results: Dual luciferase reporter gene results showed that Wnt1 is a direct target gene of miR-122 in both HepG2 and huh7 cell lines. Compared to miR-NC, anti-miR-NC, and miR-122 + Wnt1 groups, miR-122 expression was markedly higher in the miR-122 group and miR-122 + vector group, but was sharply decreased in anti-miR-122 group (both P<0.05), and the mRNA and protein levels of Wnt1, vimentin, β-cadherin, and N-cadherin decreased significantly; also E-cadherin increased, and cell proliferation, migration, and invasion decreased in the miR-122 group and miR-122 + vector group (all P<0.05), but the situation was totally reversed in the anti-miR-122 group (all P<0.05).

Conclusion: Downregulation of miR-122 promoted proliferation, migration, and invasion of human HCC cells by targeting Wnt1 and regulating Wnt/β-catenin pathway which activated the EMT pathways.

Download full-text PDF

Source
http://dx.doi.org/10.2147/OTT.S92378DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4827919PMC
April 2016
45 Reads

Publication Analysis

Top Keywords

group mir-122
16
migration invasion
12
proliferation migration
12
mir-122
12
group
9
cell lines
8
epithelial-mesenchymal transition
8
cells activating
8
activating epithelial-mesenchymal
8
e-cadherin vimentin
8
hepg2 huh7
8
vimentin β-cadherin
8
huh7 cell
8
hepatocellular carcinoma
8
β-cadherin n-cadherin
8
groups mir-122
8
invasion human
8
mir-122 vector
8
anti-mir-122 group
8
vector group
8

Similar Publications