Effect and mechanism of microRNA24 on cell proliferation and migration of osteosarcoma cell line U2OS

doi:10.3760/cma.j.issn.1673-422X.2017.07.003

Abstract

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. ResultsThe 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.

Key words: Osteosarcoma, Cell movement, MicroRNA-24

He Qihua. Effect and mechanism of microRNA24 on cell proliferation and migration of osteosarcoma cell line U2OS[J]. Journal of International Oncology, 2017, 44(7): 490-495.

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