Influence of miR -451 on biological characters of glioma cell line A172
Nan Yang, Lei Han, Anling Zhang, Peiyu Pu, Yue Zhong, Chunsheng Kang
Abstract
Nan Yang, Lei Han, Anling Zhang, Peiyu Pu, Yue Zhong, Chunsheng Kang
Abstract
Objective To investigate the influence of miR -451 on biological characters of gliomas cells A172. Method The oligonucleotide of miR -451 mimics was transfected into human glioblastoma cell line A172. The expression of miRNA -451 was identified by real - time PCR after transfection, and targeting protein expression was evaluated by Western blot. The cell proliferation was determined by flow cytometry and MTT assay. Moreover,cell cycle molecules were detected by Western blot analysis. Results The miR -451was markedly up -regulated after transfection of miRNA -451 mimics group. As compared with control and scramble group, the cell cycle was significantly arrested over 17.4% in G0/G1 phase by flow cytometry assay,meanwhile the protein expression of C - myc and regulatory molecules for cell cycle progression including CDK2, CDK4, Cyclin D1 and Cyclin E were all down -regulated. The proliferation was significantly decreased by the MTT assay at the same time. Conclusions MiR - 451 could inhibit the proliferation activity of A172 glioma cell. It suggests that miR -451 is a tumor suppressor gene in glioma. Key words: MicroRNA -451; Glioma; Cell proliferation; Cell cycle
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Objective To investigate the influence of miR -451 on biological characters of gliomas cells A172. Method The oligonucleotide of miR -451 mimics was transfected into human glioblastoma cell line A172. The expression of miRNA -451 was identified by real - time PCR after transfection, and targeting protein expression was evaluated by Western blot. The cell proliferation was determined by flow cytometry and MTT assay. Moreover,cell cycle molecules were detected by Western blot analysis. Results The miR -451was markedly up -regulated after transfection of miRNA -451 mimics group. As compared with control and scramble group, the cell cycle was significantly arrested over 17.4% in G0/G1 phase by flow cytometry assay,meanwhile the protein expression of C - myc and regulatory molecules for cell cycle progression including CDK2, CDK4, Cyclin D1 and Cyclin E were all down -regulated. The proliferation was significantly decreased by the MTT assay at the same time. Conclusions MiR - 451 could inhibit the proliferation activity of A172 glioma cell. It suggests that miR -451 is a tumor suppressor gene in glioma. Key words: MicroRNA -451; Glioma; Cell proliferation; Cell cycle
Key concepts: Cell cycle, Cell growth, Transfection, Glioma, Cyclin D1, Flow cytometry, Molecular biology, MTT assay