miR-302a對葉酸缺乏小鼠胚胎干細(xì)胞增殖和凋亡的影響
[Abstract]:Objective to investigate the effect of miR-302a on proliferation and apoptosis of embryonic stem cells (mESC) in folic acid deficient mice. Methods mESC was divided into complete medium group (control group), folic acid free medium group (folic acid free group) and folic acid free medium miR-302amimic group (miR-302a group). The expression of miR-302a in complete medium and folic acid free medium was detected by RT-PCR. MiR-302amimic, was constructed and transformed into folic acid free medium mESC. The effect of miR-302amimic on mESC activity was detected by MTT assay, the effect of miR-302amimic on apoptosis of mESC cells was detected by Annexin V-FITC/PI flow cytometry, and the effect of miR-302amimic on mESC cell cycle was detected by flow cytometry. Western blot was used to detect the activation of phosphatidylinositol 3-hydroxykinase (PI3K) / protein kinase B (Akt) / mammal rapamicin target protein (mTOR) signaling pathway, and the expression of downstream molecular cell cycle proteins D1 (CyclinD1), p21 and p27. Results RT-PCR confirmed that the expression of miR-302a in folic acid group decreased (P 0.01). Compared with the complete culture medium, the activity of mESC cells decreased, the apoptosis increased, the cell cycle arrest was in G 1 phase, the phosphorylation level of Akt and mTOR decreased, the expression of CyclinD1 was down-regulated, and the expression of p21 and p27 was up-regulated in folic acid-free medium, the difference was statistically significant (P 0.01). Compared with folic acid group, the activity of mESC cells was increased, apoptosis decreased, G 1 phase was shortened, AKT and mTOR phosphorylation level was increased, CyclinD1 expression was up-regulated, p21 and p27 expression was down-regulated in miR-302a group, the difference was statistically significant (P 0.01). Conclusion miR-302a analogues can significantly inhibit apoptosis of folic acid deficient mESC and promote its increment, which is related to PI3K/AKT/mTOR signaling pathway.
【作者單位】: 四川省醫(yī)學(xué)科學(xué)院/四川省人民醫(yī)院兒科;
【分類號】:R321
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