| 王凡 卢锦华 何杰 张浩翔 张雅鸥.MiR-147a 通过抑制细胞自噬促进HIF-1alpha蛋白的积累[J].,2017,17(2):201-204 |
| MiR-147a 通过抑制细胞自噬促进HIF-1alpha蛋白的积累 |
| MiR-147a Increases the Accumulation of HIF-1alpha Proteinthrough Inhibiting Autophagy |
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| DOI： |
| 中文关键词: 细胞自噬 miR-147a HIF-1-alpha |
| 英文关键词: Autophagy miR-147a HIF-1-alpha |
| 基金项目:深圳市科技计划项目（GJHZ20140416153718941） |
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| 中文摘要: |
| 目的：HIF-1alpha是由低氧诱导表达的一个重要的调节肿瘤生长、代谢的转录因子，它的降解除了通过泛素- 蛋白酶体途径降解
之外还与可以通过细胞自噬途径降解。通过研究miR-147a 对细胞自噬的影响从而进一步研究miR-147a 对HIF-1alpha降解的影响。
方法：在HeLa 细胞中过表达miR-147a，用Western blot 和Q-PCR 检测细胞自噬相关的标志物LC3B、P62、LAMP-2A的变化。再
通过溶酶体- 自噬泡共定位实验共聚焦显微镜观察自噬泡的数量以及共定位情况。最后通过加入自噬诱导剂（EBSS）和自噬抑制
剂（Bafilomycin A1），用Western blot 检测转染NC 与miR-147a 后HIF-1alpha蛋白的表达情况。结果：过表达miR-147a 后自噬相关的
标志物LC3B、P62 表达量上升，LAMP-2A 表达量下降，且溶酶体与自噬泡的共定位增多；加入自噬诱导剂和自噬抑制剂后
HIF-1-alpha蛋白的表达量增加。结果表明miR-147a 可以抑制细胞自噬的巨自噬途径以及分子伴侣介导的自噬途径，积累HIF-1-alpha蛋
白。结论：miR-147a通过抑制细胞自噬从而减少HIF-1-alpha蛋白的降解，但是miR-147a 作用靶点的分子机制需要进一步研究。 |
| 英文摘要: |
| Objective:HIF-1-alpha is an important hypoxia-induced factor which regulates tumor growth and metabolism. And its
degradation is associated with autophagy. To further study the impact of miR-147a on HIF-1-alpha’degradation, we study the relationship
between miR-147a and autophagy.Methods:MiR-147a was over-expressed in HeLa cells. Autophagy related markers, such as LC3B,
P62 and LAMP-2A were detected by Western blot and Q-PCR. Next, we used confocal microscope to observe the numbers of autophagic
vacuoles and its co-localization with lysosome. Finally, we performed Western blot to detect the protein level of HIF-1-alpha when HeLa cells
were transfected with miR-147a and treated with autophagy inducer or autophagy inhibitor.Results:Autophagy related markers, such as
LC3B, P62 and LAMP-2A were increased and the numbers of autophagic vacuoles and its co-localization with lysosome were raised.
The protein level of HIF-1-alpha were increased when treated with autophagy inducer or autophagy inhibitor. All these experiments revealed
that miR-147a could inhibit both macroautophagy and chaperone-mediated autophagy, thus reduced the degradation of HIF-1-alpha.Conclusion:In this paper, we found that miR-147a could inhibit autophagy and accelerated the accumulation of HIF-1-alpha through
hindering the autophagy pathway of HIF-1-alpha degradation. But the targets of miR-147a needed further research. |
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