Differential expression profile of EMT-related LncRNAs in hepatocellular carcinoma cells after incomplete radiofrequency ablation in vitro / 介入放射学杂志

ABSTRACT

Objective To investigate the differential expression profile of epithelial mesenchymal transition (EMT) related long chain noncoding RNA (LncRNA) in hepatocellular carcinoma (HCC) cells after incomplete radiofrequency ablation (RFA) in vitro.Methods Incomplete RFA was simnulated in vitro by using Huh7 cells in water bath at 47℃.EMT change was detected by microscopy and Western blot.Cell invasion and migration were detected by transwell assays.Cell proliferation was determined by cell counting kit-8 (CCK-8) assay.Differential expression profile of EMT-related LncRNAs between Huh7-H and Huh7 were analyzed by LncPath human EMT array,and it was validated by RT-PCR.Results Under microscopy,Huh7-H presented characteristic EMT morphological changes.Western blot analysis showed that the expression of E-cadherin in Huh7-H cells was decreased,while the expressions of N-cadherin and Vimentin were increased.Transwell assay test indicated that cell migration and invasion were increased in Huh7-H cell when compared with Huh7 cell (61.0±5.2 vs 138.0±11.8 and 33.3±7.8 vs 82.7±39.4,respectively),the abilities of Huh7-h cell in migration and invasion were evidently strengthened (P＜0.05).CCK-8 assay shawed that the proliferation ability of Huh 7-H was obviously higher than that Huh 7 (P＜0.05).LncPath human EMT array screened out 3 differential expressed LncRNAs (P≤0.05 and fold changes ≥ 1.5),including two down-regulated LncRNAs (FUNDC2P4,RPL27P7) and one up-regulated LncRNA (MTND4LP14).RT-PCR validation revealed that the expressions of FUNDC2P4,RPL27P7 and MTND4LP14 in Huh7-H cells were 0.137,0.869 and 1.037times of that in Huh7 cells,respectively.Clinical specimen validation showed that the expression of LncRNA FUNDC2P4 in peficancerous tissue was higher than that in HCC tissue,and the expression of LncRNA FUNDC2P4 in HCC tissue was higher than that in the residual HCC tissue after RFA.Conclusion EMTrelated LncRNA FUNDC2P4 in HCC cells with low expression after incomplete RFA is successfully screened out,which provides basis for the further investigation of the function and molecular mechanism of this gene.