| 杜 朋,杨 银,李德冠,樊飞跃,孟爱民.AFAP1在博来霉素诱导的A549细胞衰老中的作用机制研究[J].,2018,(10):1801-1805 |
| AFAP1在博来霉素诱导的A549细胞衰老中的作用机制研究 |
| The Role and Mechanism of AFAP1 in the Senescent A549 Cells Induced by Bleomycin |
| 投稿时间：2017-12-21 修订日期：2018-01-18 |
| DOI：10.13241/j.cnki.pmb.2018.10.001 |
| 中文关键词: Actin Filament Associated Protein 1(AFAP1) 衰老 A549细胞 博来霉素 |
| 英文关键词: Actin filament associated protein 1(AFAP1) Senescence A549 cell Bleomycin |
| 基金项目:国家自然科学基金面上项目(81372928，81573094) |
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| 中文摘要: |
| 摘要 目的：研究AFAP1在博来霉素诱导的A549细胞衰老模型中的作用及分子机制。方法：用50 μg/mL的博来霉素处理A549细胞5天建立细胞衰老模型。用相同浓度的博来霉素处理细胞1-5天观察细胞从周期阻滞到衰老的过程，SA-β-Gal染色检测衰老细胞数目，用Western blot方法检测AFAP1、p21、c-Src等蛋白表达。过表达AFAP1后，观察细胞衰老状态及各蛋白表达水平变化。结果：50 μg/mL的博来霉素处理A549细胞5天后可以建立细胞衰老模型，表现为BLM组SA-β-Gal阳性细胞数升高(P＜0.01)且细胞体积显著增大(P＜0.01)，p21表达水平升高。在衰老的A549细胞中，AFAP1和激活型(Src pY416)表达水平变化一致，从BLM处理后出现升高第4天开始明显下降在第5天最低，c-Src和Src pY527表达水平不变。过表达AFAP1后再用博来霉素诱导，SA-β-Gal阳性细胞数及细胞体积、Src pY416和p21表达与空载对照比较未发现有明显差异(P＞0.05)。结论：衰老的A549细胞中AFAP1表达下调，c-Src活性降低；过表达AFAP1不能减轻博来霉素诱导的A549细胞衰老，也不能抑制衰老细胞中的c-Src的活性下降。 |
| 英文摘要: |
| ABSTRACT Objective: To study the role and mechanism of AFAP1 in A549 cells senescence induced by bleomycin. Methods: A549 cells were exposed to bleomycin and then p21 and SA-β-Gal positive proportion which are markers of cellular senescence were evaluated. AFAP1 was overexpressed in A549 cells, the expression of p21, AFAP1 and c-Src were detected by western blot. Results: AFAP1 expression was decreased in senescent A549 cells, the change of which was consistent with activated c-Src (Src pY416). However, there was few variations in total c-Src as well as inhibited c-Src (Src pY527). There was no difference in the p21, Src pY416 and senes- cent phenotype between AFAP1-overexpressed cells and vehicle vector transfected control. Conclusion: AFAP1 expression decreased in senescent A549 cells induced by BLM, overexpression of AFAP1 could not alleviated the senescent phenotype and downstream molecu- lar activated c-Src. |
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