IL-37对肝癌细胞自噬的影响及作用机制的研究

发布时间：2018-07-09 13:11

本文选题：IL-37 + 自噬　；参考：《重庆医科大学》2017年硕士论文

【摘要】：目的:探讨IL-37对肝癌细胞自噬的影响及作用机制。方法:CCK-8及流式细胞术检测IL-37对肝癌细胞SMMC-7721和Huh-7增殖的影响;在电镜下观察自噬体的形态及数量;Western blot方法检测不同浓度IL-37处理的肝癌细胞中自噬蛋白LC3、Beclin1、p62,凋亡蛋白Bcl-2、Bax、Caspase3以及Akt/mTOR信号通路相关蛋白表达的差异;流式细胞术检测肝癌细胞的凋亡水平。结果:CCK-8结果显示IL-37处理组的细胞增殖率降低(p0.01);流式细胞术结果显示IL-37处理使细胞周期中G0/G1期阻滞(p0.01)和凋亡率升高(p0.05);电镜结果显示IL-37处理组自噬体的数量较对照组明显升高(p0.01);Western blot结果显示IL-37处理组中,LC3-II/LC3-I、Beclin1、Bax、Caspase3的表达均较对照组明显升高(p0.01),而p62、Bcl-2的表达明显降低(p0.01),信号通路中p-AKT、p-mTOR及相关蛋白p-p70S6K和p-4E-BP1的表达明显降低(p0.01);通过AKT激动剂IGF-1的处理,IL-37可以逆转IGF-1对LC3-II/LC3-I表达的抑制作用。结论:IL-37处理组中SMMC-7721和Huh-7细胞的增殖活性降低,自噬及凋亡率增高,且IL-37可能通过PI3K/AKT/mTOR信号通路诱导自噬的发生。本实验分析了IL-37可能通过促进肝癌细胞自噬和凋亡来降低细胞增殖,这对IL-37的进一步研究及临床靶向治疗有一定的意义。
[Abstract]:Objective: to investigate the effect and mechanism of IL-37 on autophagy of hepatoma cells. Methods the effects of IL-37 on the proliferation of SMMC-7721 and Huh-7 cells were detected by flow cytometry. The morphology and quantity of autophagy were observed under electron microscope. The expression of autophagy protein LC3 / Beclin1 / p62, apoptotic protein Bcl-2 / BaxCaspase3 and Akt / mTOR signaling pathway were detected by Western blot method in different concentrations of IL-37 treated hepatoma cells. Apoptosis level of hepatoma cells was detected by flow cytometry. Results the cell proliferation rate of IL-37 treated group was decreased (p0.01), the cell cycle arrest (p0.01) and apoptosis rate increased (p0.05) after IL-37 treatment, and the number of autophagy in IL-37 treated group was higher than that in control group (p0.05). The results of Western blot showed that the expression of Caspase3 in the IL-37 treated group was significantly higher than that in the control group (p0.01), while the expression of p62mTOR and the associated proteins p-p70S6K and p-4E-BP1 in the signal pathway were significantly decreased (p0.01), and the expression of p-p70S6K and p-4E-BP1 in the signal pathway was significantly lower than that in the control group (p0.01), while the expression of p62mTOR and the associated proteins p-p70S6K and p-4E-BP1 in the signal pathway were significantly lower than those in the control group (p0.01). IL-37 can reverse the inhibitory effect of IGF-1 on LC3-IIR-LC3-I expression. Conclusion the proliferation activity of SMMC-7721 and Huh-7 cells was decreased, the autophagy and apoptosis rate were increased, and IL-37 might induce autophagy through PI3K / AKT / mTOR signaling pathway. In this study, we analyzed that IL-37 may reduce cell proliferation by promoting autophagy and apoptosis of hepatoma cells, which may be of significance for the further study of IL-37 and its clinical targeted therapy.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.7

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