Title: Effect of miR-15b on proliferation and apoptosis of human glioma cell line A172
Abstract:Objectives
To observe the effect of miR-15b on proliferation and apoptosis of human glioma cell line A172 and to investigate its role in RAS/MEK/ERK1/2 signaling pathway cells.
Methods
The synt...Objectives
To observe the effect of miR-15b on proliferation and apoptosis of human glioma cell line A172 and to investigate its role in RAS/MEK/ERK1/2 signaling pathway cells.
Methods
The synthetic miR-15b mimics transfected the A172 cell line. Methylthiazol tetrazoliu (MTT) assay and flow cytometry were used to assess the effects of miR-15b on A172 cell proliferation and apoptosis. Western blotting assay was used to detect the effect of miR-15b on RAS and its downstream signaling pathways.
Results
After transfecting miR-15b mimics, real-time quantitative PCR was use to detect the △Ct values in the blank control group, the negative control group, the miR-15b mimics group, and they were 15.25±1.12, 15.92±0.98, and 10.58±0.87, respectively, suggesting the expression level of miR-15b in the A172 cells increased significantly. Western blot results showed that miR-15b mimics significantly inhibited the expression levels of RAS, MEK, ERK1/2, and pERK1/2 protein (all P<0.05). The test results of MTT assay showed that relative survival rates of cells were 72.28±6.52%, 61.86±5.79%, and 59.38±5.97%, respectively at 24, 48, and 72 h after the A172 cells being transfected by miR-15b mimics. The cell survival rate of the miR-15b mimics group was decreased significantly compare with the blank control group and the negative group (P<0.05). The cell cycle analysis revealed that the cell G1/G0 phase of miR-15b mimics group was 66.84%. The miR-15b mimics induced A172 cell block was at G1/G0 phase. The detection of annexin/propidium iodide (Annexin V/PI) double staining showed that the cell apoptosis rates of the blank control group, the negative control group, and the miR-15b mimics group were 2.7±0.83%, 2.5±0.81%, and 18.2±1.58%, respectively. The A172 cell apoptosis rate of the miR-15b mimics group increased significantly compare with the blank control group and the negative control group (P<0.05).
Conclusions
miR-15b can inhibit the proliferation of glioma cells and promote apoptosis. The targeted therapy of miR-15b for RAS/MEK/ERK1/2 signaling pathway is expected to become a new way for the treatment of gliomas.
Key words:
Glioma; Cell proliferation; Cell apoptosis; miR-15bRead More
Publication Year: 2015
Publication Date: 2015-11-28
Language: en
Type: article
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