白介素-27在恶性胸腔积液中的免疫调节机制

发布时间：2018-01-27 23:46

本文关键词： 白介素-27 恶性胸腔积液肺癌信号转导　出处：《华中科技大学》2015年博士论文　论文类型：学位论文

【摘要】：研究背景白介素(IL)-27是IL-12家族的新成员,大量研究认为,IL-27具有抗肿瘤细胞增殖和抗血管生成的作用。由于IL-27不仅与IL-12结构相关,它们还具有相类似的信号通路,而IL-27的毒副作用明显低于IL-12,因此,IL-27很可能代替IL-12成为新的抗肿瘤免疫制剂。然而,到目前为止IL-27在恶性胸腔积液中对肺癌细胞的免疫调节作用及其机制仍然不明确。因此,本研究主要以恶性胸腔积液为研究对象,揭示IL-27在恶性胸腔积液中的免疫调节机制。研究方法本研究收集确诊恶性胸腔积液患者的胸腔积液及外周血,通过酶联免疫吸附试验检测恶性胸腔积液上清和血清中IL-27的含量,并通过流式细胞术检测胸腔积液和外周血中分泌IL-27的CD4+T细胞的比例。同时,通过流式细胞术检测肺癌细胞表面IL-27、IFN-γ和IL-17受体的表达情况,以及经细胞因子刺激后肺癌细胞内STAT信号分子的磷酸化水平。在功能学实验中,本研究采用IL-27、IFN-γ和IL-17等刺激肺癌细胞,通过流式细胞术对比肺癌细胞增殖、凋亡和粘附情况,并通过迁移实验对比不同细胞因子对肺癌细胞迁移的影响。研究结果我们发现,IL-27在恶性胸腔积液中的含量要高于外周血,提示IL-27有可能在恶性胸腔积液中起到一定的免疫调节作用。同时,肺癌细胞表面表达较多的IL-27受体,并且通过IL-27刺激肺癌细胞,可使肺癌细胞内STAT1和STAT3信号分子磷酸化水平升高,从而激活STAT1和STAT3信号通路。对比IL-27、IFN-γ和IL-17对肺癌细胞的免疫调节作用的结果显示,与IFN-γ相类似,IL-27能抑制肺癌细胞增殖、促进肺癌细胞凋亡以及抑制肺癌细胞向胸膜间皮细胞培养上清转移,并且,IL-27抗肿瘤的效应是依赖于STAT1信号途径的,当给予STAT1抑制剂时,IL-27的抗肿瘤效应被完全逆转。不同的是,IL-27又可通过活化STAT3信号途径,发挥截然相反的作用,当给予STAT3抑制剂阻断STAT3信号途径时,IL-27的抗肿瘤的作用被增强。同时,IL-27可增加肺癌细胞和胸膜间皮细胞表面粘附分子的表达,激活STAT1信号途径可促进肺癌细胞和胸膜间皮细胞相互粘附的活性,而激活STAT3信号通路反而能够抑制这一粘附效应。这一结果提示我们,可通过激活不同的STAT信号,调节肺癌细胞与胸膜间皮细胞之间的相互粘附活性,从而达到抑制肺癌细胞向胸膜腔转移的目的。结论在恶性胸腔积液中,IL-27可通过依赖于STAT1信号途径的方式,抑制肺癌细胞增殖、促进凋亡,并且抑制肺癌细胞向胸膜腔转移,从而主要表现为抗肿瘤的效应。然而,IL-27也同时存在促进肿瘤发生发展的作用。因此,我们可通过调节IL-27活化的STAT信号通路,促进IL-27在恶性胸腔积液的微环境中发挥最大的抗肿瘤的免疫调节作用。
[Abstract]:Research background
Interleukin (IL) -27 is a new member of the IL-12 family, a large number of studies suggest that IL-27 has antiangiogenic and anti-tumor cell proliferation. Because the IL-27 is not only related with the structure of IL-12, they also have similar signaling pathways, and the toxicity of IL-27 was significantly lower than that of IL-12, therefore, IL-27 is likely to replace IL-12 a new anti tumor immune preparations. However, so far the IL-27 immunity in malignant pleural effusion of lung cancer cells and its regulation mechanism is still not clear. Therefore, this study focuses on malignant pleural effusion as the research object, to reveal the immune IL-27 in malignant pleural effusion. The method of adjusting mechanism
This study collected the diagnosis of malignant pleural effusion, pleural effusion and peripheral blood, adsorption test and serum IL-27 content in supernatant of malignant pleural effusion by ELISA, and by flow cytometry in pleural effusion and peripheral blood of secretion of IL-27 in the proportion of CD4+T cells. At the same time, through the detection of lung cancer cell surface IL-27 flow cytometry, the expression of IFN- and IL-17 gamma receptor, and lung cancer after cytokine stimulation of intracellular signaling molecules STAT phosphorylation. In functional experiment, this study used IL-27, IFN- and IL-17 gamma stimulated lung cancer cells by flow cytometry compared lung cancer cell proliferation, apoptosis and adhesion, impact and migration through experimental comparison of different cytokines on lung cancer cell migration.
Research results
We found that the content of IL-27 in malignant pleural effusion is higher than that of peripheral blood, suggesting that IL-27 may in malignant pleural effusion plays a certain role in immune regulation. At the same time, more IL-27 receptor surface expression of lung cancer cells and lung cancer cells stimulated by IL-27, STAT1 and STAT3 can increase the phosphorylation level of lung signal molecules in cancer cells, leading to activation of STAT1 and STAT3 signaling pathway. Compared to IL-27, immunoregulatory effects of IFN- gamma and IL-17 on lung cancer cells showed similar IFN- gamma, IL-27 can inhibit the proliferation of lung cancer cells and promote the apoptosis of lung cancer cells and inhibition of lung cancer cells to culture supernatant transfer, pleural mesothelial cells and anti IL-27 effect the tumor is dependent on the STAT1 signaling pathway, when treated with STAT1 inhibitors, the antitumor effect of IL-27 was completely reversed. The difference is that IL-27 can be through the activation of STAT3 signal pathway play The opposite effect, when the inhibitor of STAT3 blocking STAT3 signal pathway, IL-27 anti-tumor effect is enhanced. At the same time, IL-27 can increase the expression of lung cancer cells and mesothelial cell surface adhesion molecules, activation of STAT1 signaling pathway can promote lung cancer cells and pleural mesothelial cell adhesion activity and activation of STAT3 signaling pathway it can inhibit the adhesion effect. This result suggests that, via the activation of different STAT signals, regulating mutual adhesion activity between lung cancer cells and mesothelial cells, which can inhibit cancer cell metastasis to the pleural cavity. Conclusion
In malignant pleural effusion, IL-27 can rely on the STAT1 signaling pathway, inhibit the proliferation of lung cancer cells, promote apoptosis, and inhibit lung cancer cell metastasis to the pleural cavity, which is mainly manifested in the anti-cancer effects. However, IL-27 also promote the occurrence and development of tumor. Therefore, we can adjust the STAT signal the activation of IL-27 pathway, promote maximum antitumor immunity of IL-27 in malignant pleural effusion in the micro environment of regulation.

【学位授予单位】：华中科技大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R730.5

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