Effect of miR-129-3p targeting E2F transcription factor 5 on malignant biological behavior of hepatocellular carcinoma cells

Hua HE; Shaopeng LIU; Haichao LIU; Minghui BAI

ABSTRACT

Objective:

To investigate the effect of miR-129-3p on the proliferation, migration and invasion of hepatocellular carcinoma cells by targeting E2F transcription factor 5 (E2F5).

Methods:

The expression of miR-129-3p and E2F5 in different liver cancer cell lines and normal liver cell lines were detected by fluorescence quantitative polymerase chain reaction. HepG2 cell lines overexpressing miR-129-3p were constructed, thiazole blue cell proliferation assay and plate cloning test were used to detect the proliferation and clone formation ability of each cell. Cell scratch assay and transwell assay were used to detect cell migration and invasion. Western blotting was used to detect the protein expression. The targeting relationship between miR-129-3p and E2F5 was verified by dual luciferase reporter gene method and western blotting. The experimental groups were as follows non-transfection (NC) group; transfection control miR-con (miR-con) group; transfection of miR-129-3p (miR-129-3p) group; co-transfection of miR-129-3p and empty vector pcDNA (miR-129-3p+ pcDNA) group; co-transfection of miR-129-3p and pcDNA-E2F5 (miR-129-3p+ pcDNA-E2F5) group.

Results:

The expression of miR-129-3p in MHCC-97H, HepG2, LM3, Hep3B, PLC, HUH7 were all lower than that of HL-7702, the difference were statistically significant (all P<0.05). The number of cell clones [(86.56±20.84) vs. (511.29±45.03)and(509.78±40.81)], the cell migration rate [(3.03±1.29)% vs. (15.01±2.30)% and(14.99±2.31)%], the relative expression of migration-related protein MMP-2 [(0.51±0.22)vs.(1.87±0.30)and(1.84±0.35)]and the number of transmembrane cells [(33.10±1.58) vs. (101.23±0.31) and (100.96±3.44)] in miR-129-3p group were all decreased when compared with NC group and miR-Con group, the difference was statistically significant ( P<0.05). Dual luciferase reporting assay showed that miR-129-3p can negatively target E2F5 and inhibit its expression.

Conclusion:

The expression level of miR-129-3p is low in hepatocellular carcinoma cells, overexpression of miR-129-3p can inhibit the malignant biological behavior of hepatocellular carcinoma cells by targeting E2F5.