Effect and mechanism of microRNA-24 on cell proliferation and migration of osteosarcoma cell line U2OS
10.3760/cma.j.issn.1673-422X.2017.07.003
- VernacularTitle:微小RNA-24对骨肉瘤细胞U2OS体外增殖与迁移能力的影响及作用机制
- Author:
Qihua HE
- Keywords:
Osteosarcoma;
Cell movement;
MicroRNA-24
- From:
Journal of International Oncology
2017;44(7):490-495
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of microRNA-24 (miR-24) on cell proliferation and migration in osteosarcoma cell line U2OS and its possible mechanism.Methods U2OS cell line with miR-24 over-expression was established by transfecting miR-24 mimic, and then cell proliferation and migration in control group, negative control group and miR-24 over-expression group were detected with real-time quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) assay, cell counting kit-8 (CCK-8) assay and Transwell assay, respectively.Epithelial-mesenchymal transition (EMT) progression and activation of nuclear factor-κB (NF-κB) pathway were detected by Western blotting.Results The expressions of miR-24 in the control group, negative control group and miR-24 over-expression group were 1.00±0.00, 1.03±0.08 and 2.46±0.29, with significant difference (F=11.026, P=0.012).Compared with the control group, the expression of miR-24 in human osteosarcoma U2OS cell line was significantly increased after transfecting miR-24 mimic (t=4.604, P=0.009).After over-expression of miR-24 for 24 h, 48 h and 72 h, U2OS cell viabilities were decreased significantly compared with the control group [(3.56±0.27)% vs.(8.63±0.79)%, t=3.896, P=0.016;(20.16±1.09)% vs.(54.77±5.42)%, t=4.813, P=0.008;(45.47±3.16)% vs.(95.52±8.56)%, t=7.173, P=0.002)].After over-expression of miR-24 for 24 h, the cell migration rates in control group, negative control group and miR-24 over-expression group were (100.00±0.00)%, (99.26±5.85)% and (31.37±2.09)%, respectively, and there was statistically significant difference among the three groups (F=12.175, P=0.009);and compared with control group, cell migration rate was decreased significantly after over-expression of miR-24 (t=3.843, P=0.004).Meanwhile, over-expression of miR-24 up-regulated the expression levels of epithelial cell markers E-cadherin (t=3.852, P=0.018) and β-catenin (t=3.512, P=0.024), while down-regulated the expression levels of mesenchymal cell markers N-cadherin (t=3.832, P=0.018) and vimentin (t=4.058, P=0.012), with a suppressed EMT progress.Other than that, miR-24 over-expression inhibited the expressions of NF-κB (p65) (t=4.813, P=0.008), phosphorylation inhibitor of nuclear factor kappa-B kinase α (p-IKK-α) (t=3.764, P=0.013) and phosphorylation inhibitor of nuclear factor kappa-B kinase complex α (p-IκB-α) (t=4.064, P=0.012), suppressing the activation of NF-κB pathway.Conclusion miR-24 can suppress cell proliferation and migration of osteosarcoma cell line U2OS in vitro, and its mechanism may be related to the suppression of NF-κB activation and EMT progression.It hints that miR-24 may be used as a potential new target for osteosarcoma therapy.
