MicroRNA-206 overexpression promotes apoptosis, induces cell cycle arrest and inhibits the migration of human hepatocellular carcinoma HepG2 cells

Liu,Weiwei; Xu,Chuanming; Wan,Huifang; Liu,Chunju; Wen,Can; Lu,Hongfei; Wan,Fusheng

doi:10.3892/ijmm.2014.1800

MicroRNA-206 overexpression promotes apoptosis, induces cell cycle arrest and inhibits the migration of human hepatocellular carcinoma HepG2 cells

Authors:
- Weiwei Liu
- Chuanming Xu
- Huifang Wan
- Chunju Liu
- Can Wen
- Hongfei Lu
- Fusheng Wan
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Affiliations: Department of Biochemistry and Molecular Biology, Basic Medical College of Nanchang University, Nanchang, Jiangxi 330006, P.R. China, Medical Experiment Education Department of Nanchang University, Nanchang, Jiangxi 330006, P.R. China, Department of Clinical Laboratory, The Affiliated Hospital of Jiangxi College of Chinese Medicine, Nanchang, Jiangxi 330006, P.R. China
Published online on: June 11, 2014 https://doi.org/10.3892/ijmm.2014.1800
Pages: 420-428
Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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Abstract

MicroRNA-206 (miR-206) is known to regulate cell proliferation and migration and is involved in various types of cancer. However, the role of miR-206 in human hepatocellular carcinoma (HHC) has not been previously reported. In the present study, the expression of Notch3 in HCC and adjacent non-neoplastic tissue was immunohistochemically assessed on formalin-fixed, paraffin-embedded sections. miR-206 mimics were transiently transfected into HepG2 cells using Lipofectamine™ 2000. Subsequently, we evaluated the role of miR-206 in cell proliferation, apoptosis, cell cycle arrest and migration by MTS assay, Hoechst 33342 staining, Annexin V-FITC/PI assay, flow cytometry and wound healing assay. Using quantitative reverse transcription polymerase chain reaction (qRT‑PCR) and western blot analysis, we detected the expression of Notch3, Bax, Bcl-2, Hes1, p57 and matrix metalloproteinase (MMP)-9 at the mRNA and protein level, respectively. In addition, we measured the expression of miR-206 at the mRNA level and that of caspase-3 at the protein level. After miR-206 was upregulated in HepG2 cells, Notch3, Hes1, Bcl-2 and MMP-9 were downregulated both at the mRNA and protein level, whereas p57 and Bax were upregulated. Cleaved caspase-3 protein expression was also markedly increased. Cell proliferation was significantly attenuated and apoptosis was markedly increased. Furthermore, miR-206 overexpression induced cell cycle arrest and inhibited the migration of HepG2 cells. Taken together, our results uggest that miR-206 is a potential regulator of apoptosis, the cell cycle and migration in HepG2 cells and that it has the potential for use in the targeted therapy of HCC and is a novel tumor suppressor.

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