复发性鼻息肉分子标志物筛选及其意义

发布时间：2018-06-19 04:34

本文选题：复发性鼻息肉 + 基因芯片　；参考：《中南大学》2010年博士论文

【摘要】： 目的：应用功能分类基因芯片技术研究复发性鼻息肉组织的相关基因,并探讨其在鼻息肉发病和复发中的作用。方法：分别抽提检测10例鼻息肉组织、10例复发性鼻息肉组织、10例慢性鼻窦炎组织和10例正常鼻黏膜组织的总RNA,合成cDNA,在标准96孔PCR反应仪中进行实时定量PCR实验,用扫描仪扫描芯片荧光信号,用2-△△Ct计算JAK-STAT信号通路、MAPK信号通路和NF-κB信号通路中实验组与对照组对应基因的表达差异。为了验证这些发现的可重复性,同时也为了检验我们建立的实验体系的可靠性,我们严格按照第一次实验的设计,重复进行了第二次实验。结果：两次实验共得出与正常对照组比较上调2倍以上的差异基因14个,其中JAK-STAT信号通路9个、MAPK信号通路2个,NF-κB信号通路3个。在差异表达的基因中,主要包括信号转录与转导激活因子、转录因子、细胞因子信号转导抑制因子、表皮生长因子相关因子、细胞周期调节因子、补体因子等。其中可能反映鼻息肉复发状态的基因7个；可能反映鼻息肉状态的基因4个；可能反映鼻黏膜炎症状态的基因1个。结论：用功能分类基因芯片筛选鼻息肉、复发性鼻息肉、慢性鼻窦炎和正常鼻黏膜组织间存在的差异基因是可行的,这将为寻找鼻息肉分类和预后评估的客观实验室指标提供新的方法。目的：检测细胞周期蛋白D1(Cyclin D1)和抑瘤素M(Oncostatin M, OSM)蛋白在复发性鼻息肉、鼻息肉、慢性鼻窦炎及正常中鼻甲黏膜组织中的表达程度的差异,探讨其在复发性鼻息肉发病机制中的作用。方法：采用蛋白质印迹法分别检测6例复发性鼻息肉、6例鼻息肉、6例慢性鼻窦炎及6例正常中鼻甲黏膜组织中Cyclin D1和OSM的蛋白表达；免疫组织化学技术检测6例复发性鼻息肉、6例鼻息肉、6例慢性鼻窦炎及6例正常中鼻甲黏膜组织中Cyclin D1蛋白的表达及定位,并比较表达程度的差异。结果：(1)免疫组化显示Cyclin D1蛋白在复发性鼻息肉、鼻息肉、慢性鼻窦炎及正常中鼻甲黏膜组织中均有表达,Cyclin D1蛋白主要位于鼻黏膜上皮细胞和腺上皮细胞的细胞核,黄色或棕黄色染色为阳性。蛋白质印迹显示Cyclin D1蛋白和OSM蛋白在复发性鼻息肉、鼻息肉及慢性鼻窦炎组织中的表达均高于正常中鼻甲黏膜组织。(2)复发性鼻息肉组织中Cyclin D1和OSM蛋白表达水平高于鼻息肉、慢性鼻窦炎及中鼻甲组织,差异有统计学意义(P值均0.05)。结论：Cyclin D1和OSM在复发性鼻息肉组织中表达增高,提示它们可能在鼻息肉的复发中起着重要的作用,可作为检测鼻息肉复发的候选指标。
[Abstract]:Objective: To study the genes related to recurrent nasal polyps and to explore its role in the pathogenesis and recurrence of nasal polyps by functional classification microarray technology.
Methods: the total RNA of 10 cases of nasal polyps, 10 cases of recurrent nasal polyps, 10 cases of chronic sinusitis and 10 normal nasal mucosa were synthesized, and cDNA was synthesized. The real-time quantitative PCR experiment was carried out in the standard 96 hole PCR reaction instrument. The fluorescence signal of the chip was scanned by the scanner and the JAK-STAT signal pathway was calculated with 2- Delta Ct, and the MAPK signal was calculated. In order to verify the repeatability of these discoveries and to test the reliability of the experimental system we have established, we have repeated second experiments in strict accordance with the design of the first experiment to verify the differences in the expression of the corresponding genes between the experimental and control groups in the pathway and NF- kappa B signaling pathway.
Results: the two experiments showed that there were 14 different genes up to 2 times higher than normal controls, including 9 JAK-STAT signaling pathways, 2 MAPK signaling pathways and 3 NF- kappa B signaling pathways, including signal transcription and transduction activator, transfer factor, cytokine signal transduction inhibitor, epidermal growth factor, and epidermal growth factor. Long factor related factors, cell cycle regulating factors, complement factors, which may reflect 7 genes of recurrent nasal polyps, 4 genes that may reflect the state of nasal polyps, and 1 genes that may reflect the inflammatory state of nasal mucosa.
Conclusion: it is feasible to screen the differential genes between nasal polyps, recurrent nasal polyps, chronic rhinosinusitis and normal nasal mucosa with functional classification gene chip, which will provide a new method for finding the objective laboratory indicators for the classification of nasal polyps and the evaluation of prognosis.
Objective: to detect the difference in the expression of cyclin D1 (Cyclin D1) and tumor suppressin M (Oncostatin M, OSM) protein in recurrent nasal polyps, nasal polyps, chronic sinusitis and normal nasal mucosa, and to explore its role in the pathogenesis of recurrent nasal polyps.
Methods: the protein expression of Cyclin D1 and OSM in 6 cases of recurrent nasal polyps, 6 cases of nasal polyps, 6 cases of chronic sinusitis and 6 normal middle turbinate mucosa, 6 cases of recurrent nasal polyps, 6 cases of nasal polyps, 6 cases of slow sinusitis and 6 normal middle turbinate mucosa were detected by immunohistochemistry. The expression and location of n D1 protein and the difference of expression level were compared.
Results: (1) immunohistochemical staining showed that Cyclin D1 protein was expressed in recurrent nasal polyps, nasal polyps, chronic sinusitis and normal middle turbinate mucosa. The Cyclin D1 protein was mainly located in the nucleus of nasal epithelial cells and gland epithelial cells, yellow or brown yellow staining was positive. The Western blot showed Cyclin D1 protein and OSM protein. The expression in recurrent nasal polyps, nasal polyps and chronic sinusitis was higher than that of normal nasal mucosa. (2) the expression level of Cyclin D1 and OSM protein in recurrent nasal polyps was higher than that of nasal polyps, and the difference was statistically significant (0.05) in chronic sinusitis and middle turbinate.
Conclusion: Cyclin D1 and OSM are highly expressed in recurrent nasal polyps, suggesting that they may play an important role in the recurrence of nasal polyps, and can be used as a candidate for the detection of recurrent nasal polyps.
【学位授予单位】：中南大学
【学位级别】：博士
【学位授予年份】：2010
【分类号】：R765

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