Regulatory role of hsa-miR-214-5p on HeLa cells’ viability and apoptosis by down-regulating PAK4

Abstract:

Objective：To investigate the role of micro-RNA-214-5p（miR-241-5p） on the HeLa cells’ cell viability and apoptosis and its mechanism. Methods：HeLa cells were divided into the HeLa group，the mimic-scramble group，the miR-214 mimic group and the miR-214 inhibitor group. The miR-214 mimic and miR-214 inhibitor were used to transfect cells，the qRT-PCR was used to measure expression levels of miR-214 and P21-activated kinases 4（PAK4），the bioinformatic approach was used to predict the rela－tionship of miR-241-5p and PAK4，and the luciferase reporter assay was used to determine their relationship. The pcDNA-PAK4 （pc-PAK4），miR-214 mimic and miR-214 inhibitor were used to transfect HeLa cells，the Western blot was used to determine pro－tein level of PAK4，the CCK-8 assay was used to measure cell viability and the flow cytometry was performed to detect apoptosis. Results：The expression level of miR-214-5p in the miR-214 mimic group（46.02±5.23） was significantly higher than that in the HeLa group （1.00±0.01）（LSD-t=31.423，P=0.000）. The expression level of PAK4 in the miR-214 mimic group（0.08±0.04） was significantly lower than that in the HeLa group（0.26±0.05）（LSD-t=4.296，P=0.002）. The expression level of miR-214-5p in the miR-214 inhibitor group（0.41±0.05） was significantly lower than that in the HeLa group（1.00±0.01）（LSD-t=18.726，P=0.000）. The expression level of PAK4 in the miR-214 inhibitor group（0.58±0.05） was significantly higher than that in the HeLa group of （0.26±0.05）（LSD-t=5.061，P=0.001）. The miR-214-5p was able to be targeted-bound with PAK4. The cell growth rate in the miR-214 mimic group（3.50±0.45） was significantly lower than that in the HeLa group（5.02±0.35）（LSD-t=8.644，P=0.000）. The apoptosis rate in the miR-214 mimic group[（11.42±0.71）%] was significantly higher than that in the HeLa group[（2.61±0.63）%]（LSD-t=4.032，P=0.003）. The cell growth rate in the miR-214 inhibitor group（5.89±0.32） was significantly higher than that in the HeLa group（5.02±0.35）（LSD-t=7.863，P=0.000）. The apoptosis rate in the miR-214 inhibitor group[（0.53±0.42）%] was significantly lower than that in the HeLa group[（2.61±0.63）%]（LSD-t=4.221，P=0.002）. Co-transfection of pc-PAK4 was able to reverse the regulation function of miR-214 mimic on cell viability and apoptosis. Conclusion：The miR-214-5p inhibits the viability of HeLa cells and induces their apoptosis by down-regulating PAK4.