摘　要：本文使用茴香霉素激活神经胶质瘤U251细胞内c-jun 氨基末端激酶（JNK）活性，实现激活后的JNK对神经胶质瘤抑瘤效果的探究，并找到受JNK调节的关键转录因子。试验首先利用在线分析网站预测出受JNK调节的相关肿瘤模型，通过使用茴香霉素（JNK激动剂）实现对细胞内磷酸化JNK（P-JNK）水平的有效调节，并通过蛋白质印迹法（Western blot）探究其发挥作用的最适条件；然后通过Cell Counting Kit-8（CCK-8）检测、细胞计数、细胞划痕和流式细胞检测等试验，探究激活后的JNK对U251细胞的增殖、迁移能力以及细胞周期的影响；最后通过转录组测序（RNA-seq）技术分析找到受JNK调节的关键转录因子。试验结果表明：生物信息数据分析JNK与神经胶质瘤的发生和发展存在很强的相关性；1.0μg/mL茴香霉素作用于神经胶质瘤U251细胞1.0 h后，可有效激活细胞内源性JNK活性，抑制细胞的增殖和迁移，并导致细胞周期阻滞；RNA-seq分析揭示了JNK可能通过调节P53活性实现抑制神经胶质瘤的生长。本文研究发现，激活JNK后能够对神经胶质瘤产生抑瘤效果，为JNK可作为神经胶质瘤的治疗靶点提供了依据，通过RNA-seq分析找到的转录因子P53为进一步探究JNK影响神经胶质瘤发生发展的机制提供了方向。
关键词：JNK；神经胶质瘤细胞U251；抑瘤；转录组测序；茴香霉素

Abstract: Using anisomycin to activate JNK in glioma U251 cells, the antitumor effect of activated JNK on glioma was investigated, and the key transcription factor regulated by JNK was found. This paper uses an online analysis website to predict a tumor model regulated by JNK; through the use of anisemycin (JNK agonist), the effective regulation of intracellular P-JNK levels was achieved, and its optimal conditions are explored by Western blot experiments; through experiments such as CCK-8 detection, cell counting, cell scratching, and flow cytometry the effects of activated JNK on the cell proliferation, migration, and cell cycle of U251 cells were explored; RNA-seq analysis was used to find the key transcription factor regulated by JNK. The results show that, there is a strong correlation between JNK and the occurrence and development of gliomas through bioinformatics data analysis；after 1.0 h of 1.0μg/mL anisemycin, it can effectively activate the endogenous JNK activity of glioma U251 cells, inhibit cell proliferation and migration, and cause cell cycle arrest; RNA-seq analysis revealed that JNK may inhibit the growth of gliomas by regulating the activity of P53. This paper found that activating JNK can produce a tumor suppressive effect on gliomas, and it will provide a basis of JNK as a therapeutic target for gliomas; P53, the transcription factors found by RNA-seq analysis, that also provide a direction for further exploring the mechanism between JNK and the development of gliomas.
Key words: JNK; glioma cell U251; antitumor effect; RNA-seq; anisomycin