Suppressive effect of microRNA-451 on proliferation and apoptosis of glioma cell line A172 in vitro
Nan Yang, Yue Zhong, Chunsheng Kang, Lei Han, Anling Zhang, Peiyu Pu
Abstract
Nan Yang, Yue Zhong, Chunsheng Kang, Lei Han, Anling Zhang, Peiyu Pu
Abstract
Objective To investigate the effect of microRNA-451 (miR-451) on proliferation and apoptosis of glioma cell line A172. Methods The oligonucleotide was transfected to human glioma cell line A172. The expression of miRNA-451 was identified by Real-time polymerase chain reaction (PCR) after transfection, and targeting protein expression was evaluated by Western blotting. The cell proliferation was determined by flow cytometry and methylthiazol tetrazolium (MTT) assay. Moreover, cell proliferation and apoptosis molecules were detected by Western blotting. The number of apoptotic cells was determined by Annexin Ⅴ assay. Results After transfection of miRNA-451 mimics, as compared with control group,the miR-451 was increased larger than 6-fold in miRNA-451 mimics group, meanwhile the protein expression of AKT1 was down-regulated to (43.81 ±5.20)%, Cyclin D1 (56.09 ±3.40)% and bcl-2 (63.49± 3.70) %, conversely, p27 was up-regulated to ( 145.51 ±6.70) %. And then the cell cycle was significantly arrested in G0/G1 phase and G0/G1 phase cells were increased larger than 17.4% by flow cytometry assay. Cell proliferation was decreased larger than 35% by the MTT assay and cell apoptosis was significantly increased larger than 15% by Annexin V assay. Conclusion miR-451 could inhibit proliferation and induce apoptosis acivity of A172 glioma cells, suggesting that miR-451 was a tumor suppressor gene in glioma. Key words: microRNA-451; Glioma; Proliferation
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Objective To investigate the effect of microRNA-451 (miR-451) on proliferation and apoptosis of glioma cell line A172. Methods The oligonucleotide was transfected to human glioma cell line A172. The expression of miRNA-451 was identified by Real-time polymerase chain reaction (PCR) after transfection, and targeting protein expression was evaluated by Western blotting. The cell proliferation was determined by flow cytometry and methylthiazol tetrazolium (MTT) assay. Moreover, cell proliferation and apoptosis molecules were detected by Western blotting. The number of apoptotic cells was determined by Annexin Ⅴ assay. Results After transfection of miRNA-451 mimics, as compared with control group,the miR-451 was increased larger than 6-fold in miRNA-451 mimics group, meanwhile the protein expression of AKT1 was down-regulated to (43.81 ±5.20)%, Cyclin D1 (56.09 ±3.40)% and bcl-2 (63.49± 3.70) %, conversely, p27 was up-regulated to ( 145.51 ±6.70) %. And then the cell cycle was significantly arrested in G0/G1 phase and G0/G1 phase cells were increased larger than 17.4% by flow cytometry assay. Cell proliferation was decreased larger than 35% by the MTT assay and cell apoptosis was significantly increased larger than 15% by Annexin V assay. Conclusion miR-451 could inhibit proliferation and induce apoptosis acivity of A172 glioma cells, suggesting that miR-451 was a tumor suppressor gene in glioma. Key words: microRNA-451; Glioma; Proliferation
Key concepts: Cell growth, Apoptosis, Cell cycle, Annexin, Glioma, Molecular biology, Flow cytometry, Transfection