Effects of miR-145 on the proliferation, apoptosis and cell cycle of a human keratinocyte cell line HaCaT

Abstract

Objective To investigate the regulatory effects of miR- 145 on the proliferation, cell cycle and apoptosis of a human keratinocyte cell line HaCaT. Methods miR-145 mimics and negative control (NC) mimics were chemically synthesized and then transiently transfected into HaCaT cells respectively. After additional culture for different durations, real-time PCR was performed to determine the expression level of miR-145, MTS assay to estimate cell proliferation, and flow cytometry to detect cell apoptosis and cycle. Luciferase assay, real-time PCR and Western blot were conducted to determine whether NRAS was the target gene of miR-145. Results The miR-145 expression level in miR-145 mimic-transfected cells increased by 85.00±1.21 folds compared with NC mimic-transfected cells(t = 115.90, P 0.05). Real- time PCR and Western blot revealed that the overexpression of miR- 145 mimics had no significant effect on NRAS mRNA expression (P> 0.05), but significantly inhibited NRAS protein expression (1.52±0.07 vs. 0.20±0.02, t= 28.43, P< 0.01). Conclusion miR-145 might inhibit proliferation and promote apoptosis of HaCaT cells by influencing cell cycle via NRAS. Key words: Keratinocytes; Cell proliferation; Cell cycle; Apoptosis; Condylomata acuminata; HaCaT cells; miR-145; NRAS