表皮生长因子刺激对肺腺癌细胞系产生的外泌体的影响

发布时间：2018-05-31 22:38

本文选题：表皮生长因子 + 外泌体　；参考：《北京协和医学院》2016年博士论文

【摘要】：背景肺癌是我国乃至全世界发病率和死亡率最高的肿瘤,其进展快、恶性程度高且极其容易转移。肿瘤产生的外泌体在肿瘤的发生、侵袭和转移方面都扮演者重要角色。表皮生长因子受体(Epidermal Growth Factor Receptor, EGFR)的过表达与EGFR信号通路的激活与肺腺癌的增殖、侵袭、转移、密切相关。但关于EGFR通路激活对肺腺癌产生的外泌体的影响,目前研究尚不够。目的观察表皮生长因子(Epidermal Growth Factor, EGF)刺激对肺腺癌细胞系产生的外泌体形态、粒径以及分泌量的影响,并探讨EGFR相关的促肿瘤增殖、转移作用能否能通过外泌体在肿瘤细胞间传递。方法使用EGF刺激人肺腺癌H322细胞,梯度离心法分离其细胞上清中外泌体。采用透射电镜、纳米示踪分析、Western蛋白印记以及外泌体蛋白定量方法鉴定并比较EGF刺激组与对照组所得外泌体在形态、粒径以及分泌量方面的区别。将上述两组外泌体与肺腺癌细胞共孵育后,检测肺腺癌细胞在增殖和转移方面的变化。结果梯度离心所得到的颗粒具有外泌体典型杯状的结构,颗粒直径主要分布在40-150nm之间,表达外泌体标志性蛋白CD63和TSG101。EGF处理组与对照组相比,肺腺癌产生的外泌体在形态、粒径分布无明显影响,但分泌量明显增加。两组外泌体都可促进肺腺癌细胞的增殖,但组间无统计学差异差异。而EGF处理组外泌体较对照组能提高肺腺癌的迁移能力,且具有统计学意义。结论EGF刺激对肺腺癌细胞产生的外泌体的形态和粒径影响不明显,但可促进外泌体的分泌。EGFR相关促肿瘤转移作用可能通过外泌体在肿瘤细胞间传递。
[Abstract]:Background Lung cancer is a tumor with the highest morbidity and mortality in China and the world. Tumor-producing exocrine plays an important role in tumorigenesis, invasion and metastasis. Overexpression of epidermal Growth Factor Receptor, EGFR) and activation of EGFR signaling pathway are closely related to proliferation, invasion and metastasis of lung adenocarcinoma. However, the effect of EGFR pathway activation on exocrine secretion in lung adenocarcinoma is not well studied. Objective to observe the effects of epidermal Growth Factor, EGF) stimulation on the morphology, size and secretion of exocrine in lung adenocarcinoma cell lines, and to explore the role of EGFR in promoting tumor proliferation and whether metastasis can be transmitted between tumor cells through exocrine. Methods Human lung adenocarcinoma cell line H 322 was stimulated by EGF and its supernatant was isolated by gradient centrifugation. Transmission electron microscopy, nano-tracer analysis and quantitative analysis of Western protein imprinting and exocrine protein were used to identify and compare the morphology, particle size and secretion of exocrine in EGF stimulated group and control group. After the two groups were incubated with lung adenocarcinoma cells, the proliferation and metastasis of lung adenocarcinoma cells were detected. Results the granules obtained by gradient centrifugation had the typical goblet structure of exocrine, and the diameter of the granules mainly distributed between 40-150nm. The exocrine bodies produced by lung adenocarcinoma were in shape in the treatment group of CD63 and TSG101.EGF, which expressed the hallmark protein of exocrine, compared with the control group. The particle size distribution had no obvious effect, but the secretion increased obviously. Both exocrine bodies could promote the proliferation of lung adenocarcinoma cells, but there was no statistical difference between the two groups. The exocrine body in EGF group could improve the migration ability of lung adenocarcinoma compared with the control group, and had statistical significance. Conclusion EGF stimulation has no significant effect on the morphology and particle size of exocrine bodies produced by lung adenocarcinoma cells, but it can promote the secretion of exocrine. EGFR-related tumor metastasis may be transmitted between tumor cells through exosomes.
【学位授予单位】：北京协和医学院
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R734.2

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