RESUMO
Objective To explore the effect of invasion and migration of hepatocellular carcinoma (HCC) endothelial cells (TECs) affected by overexpression of microRNA-3178 (miR-3178) through the transfection of miR-3178 mimic.Methods Real-time polymerase chain reaction (Real-time PCR) was used to identify differential expression of miR-3178 in normal hepatic sinusoidal endothelial cells (HSECs) and HCC TECs.Furthermore,HCC TECs were divided into 3 groups:control (CON) group,miRNA-3178 upregulation (Mimics,up-regulation of miR-3178 expression was achieved using miR-3178 mimics transfected into HCC TECs) group and negative control (NC,negative control sequence was transfected into HCC TECs) group.RT-PCR was used to detect expression of miR-3178 in HCC TECs before and after transfection.Transfection efficiency was observed by using an inverted fluorescence microscope.HCC TECs invasionand migration were measured by matrigel invasion and transwell migration assay.EGR3 protein expression of HCC TECs were identified by Western blotting analysis.EGR3 mRNA expression of HCC TECs were identified by RT-PCR analysis.Results The results of RT-PCR showed that miR-3178 was significantly down-regulated in HCC TECs compared to HSECs (P ＜0.05),and expression of miR-3178 was significantly increased after the transcienttransfection (P ＜ 0.05).The transfection efficiency in HCC TECs was morethan 90％.Number of migrated and invaded cells and in miR-3178 group was significantly less than those in other groups.Target gene prediction software showed EGR3 was a possible candidate target.Transfection of miR-3178 mimic significantly decreased the mRNA and protein expression levels of EGR3.Conclusion MiR-3178 was downregulated in HCC TECs and overexpression of miR-3178 can specifically inhibit migration and invasion of HCC TECsin vitro through inhibiting EGR3 expression,thus,miR-3178 might be a critical targeted therapy strategv for HCC.
RESUMO
Objective To study the relationship between the expression level of human telomerase reverse transcriptase (hTERT) protein and telomerase activity as well as its clinical significance in hepatocellular carcinoma (HCC). Methods Immunohistochemical SP method was employed to detect hTERT protein expression in 52 cases of HCC and paired adjacent tumor tissues, and telomerase activity was examined in these samples with telomeric repeat amplification-ELISA. Results The positive rates of hTERT protein expression and telomerase activity in HCC were 86.5%(45/52) and 80.8%(42/52), respectively, which were significantly higher compared with the adjacent tumor tissues(P