MicroRNA-206 overexpression promotes apoptosis, induces cell cycle arrest and inhibits the migration of human hepatocellular carcinoma HepG2 cells
| Autor: | Huifang Wan, Chuanming Xu, Weiwei Liu, Chunju Liu, Can Wen, Hongfei Lu, Fusheng Wan |
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| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
Cell cycle checkpoint
Carcinoma Hepatocellular tumor suppressor Biology migration Flow cytometry Annexin Cell Movement microRNA Genetics medicine Humans Genes Tumor Suppressor medicine.diagnostic_test Cell growth Liver Neoplasms apoptosis General Medicine Articles hepatocellular carcinoma Cell Cycle Checkpoints Hep G2 Cells Cell cycle Molecular biology Neoplasm Proteins Reverse transcription polymerase chain reaction Gene Expression Regulation Neoplastic MicroRNAs Apoptosis cell cycle arrest microRNA-206 |
| Zdroj: | International Journal of Molecular Medicine |
| ISSN: | 1791-244X 1107-3756 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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