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- 2016
Eca109细胞中自噬模型的建立
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Abstract:
摘要 目的:在食管鳞癌细胞Eca109中建立自噬模型. 方法:按照常规细胞培养方法,将pmcherry-GFP-LC3质粒转染进入Eca109细胞, mcherry为红色荧光蛋白,GFP为绿色荧光蛋白,可示踪自噬体形成.加入浓度为50 nmol/L自噬激活剂雷帕霉素,建立自噬模型.采用电镜观察雷帕霉素诱导后细胞中自噬小体的产生,激光共聚焦显微镜观察自噬发展的阶段,并对发生自噬行为的细胞进行计数.结果: 雷帕霉素诱导后可见明显的自噬小体,激光共聚焦采图可见自噬细胞数量明显增多(P<0.05).结论: 1.在Eca109细胞中通过加入自噬诱导剂雷帕霉素成功构建细胞自噬模型;2.细胞自噬模型的建立为研究肿瘤细胞自噬活动的现象和分子调控机制提供了新的实验平台.
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