纳米二氧化硅刺激BEAS-2B细胞炎症和纤维化反应及其与外周血相关细胞因子的关系研究

发布时间：2018-01-06 01:33

本文关键词：纳米二氧化硅刺激BEAS-2B细胞炎症和纤维化反应及其与外周血相关细胞因子的关系研究　出处：《昆明医科大学》2015年硕士论文　论文类型：学位论文

【摘要】：背景与目的：肺癌是发生于支气管粘膜上皮的恶性肿瘤。肺癌的病因至今尚不明确。云南省是我国肺癌的高发区,特别是宣威地区的女性肺癌发病率居世界首位,前期相关研究显示宣威地区肺癌的高发可能与燃煤中含有大量的纳米二氧化硅颗粒,导致人吸入烟尘后肺部出现炎症反应有关。参与肺部炎症反应的细胞因子可分为促肿瘤类因子：IL-6、IL-8、IL-17、RANTES和MIP-1a;抗肿瘤类因子：IL-4、IL-13、IL-2、TNF-a, IFN-γ;促进血管生成及纤维化类因子：VEGF、b-FGF, MCP-1、IL-12、IL-5。还有比较特殊的有IL-10,其对肿瘤的发生发展具有两面性,既有利于肿瘤的生存又能抑制肿瘤生长。本研究探讨纳米二氧化硅与人正常支气管上皮细胞(BEAS-2B细胞)的关系,探索炎症反应在肺癌发生和进展中的作用,为肿瘤寻找合适的标记物奠定基础。方法：1、分组采用体外细胞培养的方式,在1640培养基中加入不同粒径纳米二氧化硅,通过不同粒径纳米二氧化硅刺激人正常支气管上皮细胞(BEAS-2B细胞),并于24h后观察其形态学大小、细胞结构的改变；浓度为10μg/mL、25μg/mL、50ug/mL进行实验,一次性刺激24h后,经过离心后取上层清液。2、采用Bio-Plex悬液芯片系统检测纳米二氧化硅刺激BEAS-2B细胞24h后培养上清液中促肿瘤类因子：IL-6、IL-8、IL-17、RANTES和MIP-1a;抗肿瘤类因子：IL-4、IL-13、IL-2、TNF-a、IFN-γ;促进血管生成及纤维化类因子：VEGF、 b-FGF,MCP-1、IL-12、IL-5;同时具有促肿瘤及抗肿瘤作用炎性因子IL-10的水平变化。3、收集宣威地区女性非小细胞肺癌(NSCLC)患者、非宣威地区女性非小细胞肺癌患者外周血各20例、健康女性的外周血20例,按照不同临床分期、病理类型、分化程度、淋巴结转移进行统计,经过离心后取上层血清。采用Bio-Plex悬液芯片系统检测血清中促肿瘤了类因子:IL-6、IL-8、IL-17、RANTES和MIP-1a;抗肿瘤类因子：IL-4、IL-13、IL-2、TNF-a、IFN-γ:促进血管生成及纤维化类因子：VEGF、b-FGF, MCP-1、IL-12、IL-5:同时具有促肿瘤及抗肿瘤作用炎性因子IL-10的水平变化。结果：1、纳米二氧化硅刺激下BEAS-2B细胞形态学改变：纳米二氧化硅刺激细胞24h后,与对照组相比,在镜下观察可见实验组细胞生长状态欠佳,细胞体积增大,部分细胞胞质内可见空泡样改变,细胞边缘变钝、不规则,50ug/mL浓度下部分细胞出现漂浮。2、纳米二氧化硅致BEAS-2B细胞炎性反应：各浓度刺激下,不同粒径的纳米二氧化硅可引起BEAS-2B细胞上清液中抗肿瘤类因子：RANTES因子在30nm组与50nm组之间存在差异性(P0.05)。促进血管生成及纤维化类因子：、EGF因子在对照组与30nm组之间存在差异性(P0.05),IL-12因子在对照组与30nm、 50nm组之间存在差异性(P0.05)；同时具有促肿瘤及抗肿瘤作用炎性因子IL-10在对照组、30nm组、50nm组各组之间均存在差异性(P0.05)。3、宣威地区女性肺癌外周血：宣威地区女性肺癌患者、非宣威地区女性肺癌患者以及健康女性血清中抗肿瘤类因子：IL-6因子在正常组与宣威组之间存在显著差异性(P0.01),IL17因子在正常组与宣威组、非宣威组之间存在差异性(P0.05)。抗肿瘤类因子：IL-4因子在正常组与非宣威组之间存在差异性(P0.05),IL-13在正常组与宣威组、非宣威组之间存在显著差异性(P0.01),IFN-γ在正常组与宣威组之间存在差异性(P0.05);促进血管生成及纤维化类因子：VEGF因子在正常组与宣威组、非宣威组之间存在显著差异性(P0.01),b-FGF因子在正常组与非宣威组之间存在差异性(P0.05),IL-12因子在正常组与宣威组、非宣威组之间存在显著差异性(P0.01)；同时具有促肿瘤及抗肿瘤作用炎性因子IL-10在正常组与宣威组之间存在差异性(P0.05)、在正常组与非宣威组之间存在显著差异性(P0.01)。结论：1、纳米二氧化硅对人支气管上皮细胞有刺激作用,能促进支气管上皮发生重度不典型增生,可能与肺癌发生有关。2、通过检测炎症相关及纤维化相关细胞因子的分泌状态可在一定程度上提示患者病情变化。3、宣威女性肺癌患者外周血存在差异性表达的细胞因子,可能与云南宣威女性肺癌的高发原因有关。
[Abstract]:Background and objective: lung cancer is occurring in bronchial epithelial malignant tumors. The etiology of lung cancer is still not clear. China's Yunnan province is a high incidence of lung cancer, especially in female lung cancer Xuanwei rate ranks first in the world, the research shows that Xuanwei lung cancer may contain a large number of nano silica particles and coal in the cause of inflammation of lung after smoke inhalation. Those cytokines involved in lung inflammation can be divided into types of tumor promoting factors: IL-6, IL-8, IL-17, RANTES and MIP-1a; antitumor factor: IL-4, IL-13, IL-2, TNF-a, IFN- gamma; promote angiogenesis and fibrosis cytokines: VEGF. B-FGF, MCP-1, IL-12, IL-5. and the special IL-10, with the two sides of the tumor development, is conducive to the survival of tumor and inhibit tumor growth. In this study, two nano oxide The silicon and the normal human bronchial epithelial cells (BEAS-2B cells) to explore the relationship of inflammation in lung cancer and its role in the progress, lay the foundation for appropriate markers for tumor. Methods: 1 groups using cell culture in vitro, in 1640 medium was added with particle size of nano silica by different particle the diameter of nano silica stimulation of normal human bronchial epithelial cells (BEAS-2B cells), and observe the morphology size after 24h, the change of cell structure; the concentration of 10 g/mL, 25 g/mL, 50ug/mL experiment, one-time after 24h stimulation, after centrifugation the supernatant liquid of.2, using Bio-Plex chip detection system of nano silica suspension stimulation of BEAS-2B cells after cultured 24h tumor promoting factor was: IL-6, IL-8, IL-17, RANTES and MIP-1a; antitumor factor: IL-4, IL-13, IL-2, TNF-a, IFN- gamma; promote angiogenesis and fiber Classes: VEGF, b-FGF, factor MCP-1, IL-12, IL-5;.3 levels also has tumor promoting and antitumor effects of inflammatory factor IL-10, collected the Xuanwei female non-small cell lung cancer (NSCLC) patients, female non Xuanwei patients with non-small cell lung cancer in peripheral blood of the 20 cases, 20 cases of peripheral the blood of healthy women, according to the different clinical stage, pathological type, differentiation degree, lymph node metastasis were analyzed by centrifugation after taking the serum. Using Bio-Plex suspension array system for detection of serum tumor promoting factor class: IL-6, IL-8, IL-17, RANTES and MIP-1a; antitumor factor: IL-4, IL-13, IL-2 TNF-a, IFN-, Gamma: promote angiogenesis and fibrosis cytokines: VEGF, b-FGF, MCP-1, IL-12, IL-5: levels also can promote tumor and anti-tumor effect of inflammatory factor IL-10. Results: 1, nano silica thorn morphology of BEAS-2B cell change: two nm excitation The silicon oxide stimulation of 24h cells, compared with control group, experimental group was observed under the microscope the growth state of poor cell volume increases, some cells in cytoplasm vacuolization, cell edge blunt, irregular, floating part of.2 cell 50ug/mL concentration, nano silica induced BEAS-2B cell inflammatory response the concentration under the stimulation of silica nanoparticles with different diameters can be induced by anti-tumor cytokines in the supernatants of BEAS-2B cells: RANTES factor differences between 30nm group and 50nm group (P0.05). To promote the angiogenesis and fibrosis factor: EGF factor in the difference between the control group and 30nm group (P0.05). The IL-12 factor in the control group and 30nm, the differences between 50nm groups (P0.05); can promote tumor and anti-tumor effect of inflammatory factor IL-10 in the control group, 30nm group at the same time, there are differences between groups 50nm (P0.05).3 The Xuanwei female lung cancer, peripheral blood: Xuanwei female lung cancer patients, tumor factors of female lung cancer patients in Xuanwei area and non healthy women in serum: factor IL-6 had significant differences between the normal group and Xuanwei group (P0.01), IL17 factor in normal group and Xuanwei group, there are differences between non Xuanwei group (P0.05). The antitumor factor: IL-4 factor differences between normal group and non Xuanwei group (P0.05), IL-13 in normal group and Xuanwei group, there were significant differences between non Xuanwei group (P0.01), IFN- gamma differences between normal group and Xuanwei group (P0.05); angiogenesis the formation and fibrosis factor: factor VEGF in normal group and Xuanwei group, there were significant differences between non Xuanwei group (P0.01), b-FGF factor differences between normal group and non Xuanwei group (P0.05), IL-12 factor in normal group and non Xuanwei group. There is a significant difference between Xuanwei group (P0.01); at the same time can promote tumor and anti-tumor effect of inflammatory factor IL-10 differences between normal group and Xuanwei group (P0.05), there is a significant difference between normal group and non Xuanwei group (P0.01). Conclusion: 1. The nano silicon oxide has a stimulating effect two on human bronchial epithelial cells, can promote the occurrence of severe bronchial epithelial dysplasia, may occur on.2 and lung cancer, by detecting the secretion of inflammatory state and fibrosis related cytokines to a certain extent, suggesting that changes in patients with.3, patients of Xuanwei female lung cancer peripheral blood cytokines are differentially expressed, may cause high incidence Yunnan Xuanwei female lung cancer.

【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R734.2

【参考文献】