miR-216b Inhibits Cervical Cancer Cells Autophagy by Regulating Beclin-1 Expression
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摘要:目的
研究miR-216b影响宫颈癌HeLa细胞自噬的作用机制。
方法转染miR-216b及应用其抑制剂后，利用GFP-LC3 shRNA转染等方法检测HeLa细胞的自噬水平，Western blot检测自噬相关基因Beclin-1和LC3-Ⅱ的表达变化。
结果HeLa细胞转染miR-216b后，HeLa细胞中自噬相关基因Beclin-1受到抑制，LC3-Ⅱ的表达增加，细胞自噬受抑，而抑制miR-216b的表达，自噬水平升高。进一步研究发现，miR-216b能够通过靶结合Beclin-1 3’UTR抑制Beclin-1表达从而抑制细胞自噬的发生。
结论miR-216b能够抑制宫颈癌HeLa细胞发生自噬，为宫颈癌的临床治疗提供了的理论基础。
Abstract:ObjectiveTo investigate the effect of miR-216b on autophagy in cervical cancer HeLa cells.
MethodsAfter the transfection of miR-216b and the application of their inhibitors,we used GFP-LC3 shRNA transfection to test the autophagy levels of HeLa cells and Western blot to test the expression changes of autophagy-related genes Beclin-1 and LC3-Ⅱ.
ResultsAfter transfected with miR-216b,autophagy related gene Beclin-1 was inhibited in HeLa cells,the expression of LC3-Ⅱ was increased,and autophagy was inhibited; however,the level of autophagy was increased when the expression of miR-216b was inhibited. Further,miR-216b can inhibit the occurrence of autophagy by inhibiting the expression of Beclin-1 by targeting Beclin-1 3’UTR.
ConclusionmiR-216b could inhibit HeLa cells autophagy,which provides a theoretical basis for the clinical treatment of cervical cancer.
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Key words:
- miR-216b /
- Cervical cancer /
- Autophagy /
- Beclin-1
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图 1 荧光显微镜观察经miR-216b转染或雷帕霉素处理后HeLa细胞中GFP-LC3自噬点的分布变化
Figure 1 Distribution of GFP-LC3 autophagy points in HeLa cells after transfection of miR-216b or rapamycin treatment observed by fluorescence microscopy
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图 2 Western blot检测miR-216b抑制细胞自噬水平
Figure 2 Cell autophagy inhibited by miR-216b detected by Western blot
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图 3 荧光显微镜观察经miR-216b抑制剂转染处理后HeLa中GFP-LC3自噬点的分布变化
Figure 3 Distribution of GFP-LC3 autophagy points in HeLa cells after transfection with miR-216b inhibitor observed by fluorescence microscopy
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图 4 Western blot检测发现抑制miR-216b表达能够促进细胞自噬水平
Figure 4 Inhibiting miR-216b expression could promote cells autophagy level observed by Western blot
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图 5 数据库分析miR-216b能够靶结合Beclin-1 3’UTR序列
Figure 5 miR-216b could targetedly binding Beclin-1 3'UTR sequence analyzed by database
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