MiR-429 regulates the proliferation and apoptosis of nephroblastoma cells through targeting c-myc.

OBJECTIVE

We explored the regulation of miR-429 in nephroblastoma cells, investigating the mechanisms by which miR-429 influenced the proliferation and apoptosis of nephroblastoma.

PATIENTS AND METHODS

The quantitative reverse transcription polymerase chain reaction (qRT-PCR) was conducted to detect the expression of miR-429 in nephroblastoma tissues and cell lines (G401). The interaction between miR-429 and c-myc was confirmed by qRT-PCR, Western blotting and luciferase assays, respectively. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) and clone formation assay were used to detect the effect of miR-429 on the proliferation and clone formation ability of G401 cells. Cell cycle and apoptosis of G401 cells were measured by flow cytometry and TUNEL staining, respectively.

RESULTS

miR-429 was down-regulated in nephroblastoma tissues and cells. On-line target gene prediction software was applied to screen c-myc, the potential downstream target gene of miR-429. The expression of c-myc was negatively regulated by miR-429. Subsequent experiments demonstrated that overexpression of miR-429 inhibited the proliferation ability and arrested cell cycle of G401 cells in G0/G1 phase. Besides, the number of apoptotic cells was increased in miR-429 intervened group. However, c-myc could reverse the biological effects of miR-429 on proliferation, apoptosis, clone formation and cell cycle of G401 cells.

CONCLUSIONS

MiR-429 can regulate the proliferation, clone formation, cell cycle and apoptosis of nephroblastoma cells through targeting c-myc, indicating miR-429 may function as a potential therapeutic target for the treatment of nephroblastoma.