口腔扁平苔藓发病机制初探及癌前监测手段评估

发布时间：2018-03-03 07:16

本文选题：口腔扁平苔藓　切入点：外周血淋巴细胞　出处：《南京医科大学》2014年硕士论文　论文类型：学位论文

【摘要】：目的：探索口腔扁平苔藓(Oral Lichen Planus, OLP)外周血淋巴细胞中游离钙[Ca2+]i水平,及与之密切相关的两种调控蛋白：Golli-MBP (Genes of the Oligodendrocyte lineage-Myelin Basic Protein,少突胶质细胞谱系基因-髓鞘碱性蛋白)和STIM1(Stromal Interaction Molecule1,基质相互作用分子1)的表达水平,探讨Golli-MBP、STM1对口腔扁平苔藓外周免疫细胞的影响,初步探索口腔扁平苔藓的发病机制。方法：本实验采用钙离子荧光探针Fluo-3检测43例OLP患者(22例非糜烂型及21例糜烂型)外周血淋巴细胞和20例正常人外周血淋巴细胞中游离钙离子水平。采用实时定量PCR(Quantitive Real-Time PCR, qPCR)和免疫蛋白印记技术(Western Blot),分别检测43例OLP患者和20例正常人外周血淋巴细胞中Golli-MBP和STIM1的基因和蛋白表达水平。统计学分析各组数据,比较各组指标是否存在统计学差异。结果：检测结果表明,OLP患者外周血淋巴细胞受到活化刺激后,胞内游离钙离子升高幅度显著低于正常对照组(P0.001)。Golli-MBP mRNA和蛋白在OLP组中的表达显著高于正常对照组(P0.001)。同样,STIM1mRNA和蛋白在OLP组中的表达亦显著高于正常对照组(P0.01)。结论：OLP患者外周血淋巴细胞胞内游离钙离子浓度在活化过程中升高水平低于对照组,提示OLP的淋巴细胞钙离子信号通路可能存在异常,该通路涉及的外周血免疫细胞活化可能存在障碍。本实验亦发现Golli-MBP和STIM1在OLP患者外周血淋巴细胞中高表达,可能导致机体外周免疫系统失衡,这可能与OLP的病程迁延、反复发作有一定关系。日的：评估脱落细胞DNA含量分析法监测口腔黏膜潜在恶性病变癌变的灵敏度、特异度及其实用性,寻求更有效的口腔黏膜癌前诊断的无创性监测手段。方法：本实验采用细胞刷收集了52例口腔黏膜潜在恶性病变患者的口腔黏膜病损处脱落细胞,同时切取相同部位进行病理活检。用全自动DNA含量检测仪检测口腔黏膜脱落细胞的DNA含量,出现细胞DNA含量大于5c,诊断为阳性。以病理活检报告作为诊断金标准,评估脱落细胞DNA含量检测的敏感度和精确度。结果：以病理活检报告作为诊断金标准,脱落细胞DNA倍体分析的灵敏度为86.36%,特异度为90.00%,假阳性率为13.64%,假阴性率为10.00%,阳性预测价值为86.36%,阴性预测价值为90.00%。结论：脱落细胞DNA含量检测作为一种无创性监测手段,在对口腔黏膜潜在恶性病变癌变监测中具有较高灵敏度和特异度,患者依从性高,适于作为一种癌前监测手段,监测口腔黏膜潜在恶性病变病情进展状况。
[Abstract]:Objective: to explore the level of free calcium [Ca2] I in peripheral blood lymphocytes of oral lichen planus (OLP). And the expression levels of two closely related proteins: Golli-MBP genes of the Oligodendrocyte lineage-Myelin Basic protein (oligodendrocyte lineage gene-myelin basic protein) and STIM1(Stromal Interaction Molecule1, matrix interaction molecule 1). To investigate the effect of Golli-MBP STM1 on peripheral immune cells in oral lichen planus and to explore the pathogenesis of oral lichen planus. Methods: calcium fluorescence probe Fluo-3 was used to detect the level of free calcium ion in peripheral blood lymphocytes of 22 cases of non-erosive type and 21 cases of erosive type in 43 cases of OLP and 20 cases of normal people. The gene and protein expression of Golli-MBP and STIM1 in peripheral blood lymphocytes of 43 OLP patients and 20 normal controls were detected by quantitative PCR(Quantitive Real-Time PCR (qPCR) and immuno-protein imprinting technique. To compare whether there were statistical differences in the indexes of each group. Results: the results showed that the peripheral blood lymphocytes of OLP patients were stimulated by activation. The expression of intracellular free calcium mRNA and protein in OLP group was significantly higher than that in normal control group (P 0.001N. Golli-MBP mRNA and protein), and the expression of STIM1 mRNA and protein in OLP group was also significantly higher than that in normal control group (P 0.01). Conclusion the level of intracellular free calcium ion in peripheral blood lymphocytes of patients with OLP is lower than that of control group during activation, suggesting that the calcium signaling pathway of lymphocytes in OLP may be abnormal. This study also found that the high expression of Golli-MBP and STIM1 in peripheral blood lymphocytes of OLP patients may lead to the imbalance of peripheral immune system, which may delay the course of OLP. Repeated attacks have something to do with it. Objective: to evaluate the sensitivity, specificity and practicability of exfoliated cell DNA assay in the detection of potential malignant lesions of oral mucosa, and to seek a more effective non-invasive monitoring method for precancerous diagnosis of oral mucosa. Methods: the abscission cells were collected from 52 patients with potential malignant lesions of oral mucosa by cell brush. At the same time, the same site was removed for pathological biopsy. The DNA content of the exfoliated cells of oral mucosa was detected by automatic DNA content detector. The DNA content of the cells was more than 5c. the diagnosis was positive. The pathological biopsy report was used as the diagnostic gold standard. To evaluate the sensitivity and accuracy of DNA content detection in exfoliated cells. Results: with pathological biopsy report as diagnostic gold standard, the sensitivity, specificity, false positive rate, false negative rate, positive predictive value and negative predictive value of DNA ploidy analysis in exfoliated cells were 86.36, 90.00,13.64, 10.00, respectively, respectively. Conclusion: as a noninvasive monitoring method, the detection of DNA content in exfoliated cells has high sensitivity and specificity in the monitoring of potential malignant lesions of oral mucosa, and the patient's compliance is high, so it is suitable for precancerous monitoring. To monitor the progress of oral mucosal potential malignant lesions.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R781.5

【共引文献】