血清特异性microRNA作为尘肺早期诊断生物标志物的研究

发布时间：2018-03-14 18:32

本文选题：尘肺　切入点：miRNA　出处：《天津医科大学》2013年硕士论文　论文类型：学位论文

【摘要】：目的目前,对尘肺的诊断主要采取职业史结合影像学的方式,一旦确诊,病人肺纤维化进程已无法逆转。虽然尘肺病的防治主要在于控制工作环境的粉尘浓度,但是探索早期的诊断指标,从“二级预防”出发,为预防尘肺提供科学依据。因此,寻找尘肺早期诊断的生物标志物,对于早期发现疑似病人,早治疗,降低尘肺和职业性肿瘤等重大职业病的发病率,具有重大的经济和社会效益。本课题旨在筛选尘肺病人血清中特异表达的microRNA(miRNA),探索血清miRNA作为尘肺早期生物标志物的诊断价值。方法采用TaqMan低密度芯片(TLDA)的方法,筛选出候选的可作为尘肺的筛查诊断及预后预测的血清miRNA标志物,并采用RT-qPCR方法进行验证。通过病例对照研究和统计分析,得到可用于尘肺筛查诊断的血清生物标志物,并绘制受试者工作曲线评估候选的血清miRNA标志物对尘肺的诊断价值,比较其灵敏度和特异性。最终拟合出多个候选miRNA用于尘肺早期筛查、早期诊断、预后预测的联合应用模型。结果 1.通过TaqMan低密度芯片方法筛选出在正常对照组、尘肺组之间血清中有一定表达差异的10个候选miRNA,用RT-qPCR方法在Ⅰ期尘肺130例,Ⅱ期尘肺22例和对照152例血清样本中进行验证,均可检测到有效表达。 2.与健康对照组相比,尘肺样本中6个miRNA呈高表达,分别为miR-21、 miR-200c、miR-16、miR-206、miR-155、miR-29a,1个呈低表达(miR-204),差异均有显著性。 3. Ⅰ、Ⅱ期尘肺间的比较：结果显示miR-204、miR-21在Ⅰ、Ⅱ期尘肺中的表达量存在差异性,miR-21在对照组以及Ⅰ、Ⅱ期尘肺中表达递增,miR-204在对照组以及Ⅰ、Ⅱ期尘肺中表达递减。 4.拟合6个有尘肺筛查诊断价值且灵敏度和特异性良好的血清miRNA建立尘肺和正常人群中筛查诊断的Logistic回归模型：logitP=13.769+0.536×miR-21—0.878×miR-200C—0.012×miR-16—0.111×miR-206+0.117×miR-155—1.192×miR-29a,其AUC-ROC为0.980(95%CI:0.970-0.993, P＜0.0001)。结论 1.利用TaqMan低密度芯片绘制尘肺血清miRNA表达谱,方法可靠可行。 2.血清miRNA可望作为尘肺筛查诊断的标志物,具有潜在的应用价值。 3.通过血清miRNA的表达水平,可以提示尘肺发病进程。 4.多个血清miRNA联合应用检测可改善尘肺诊断的灵敏度和特异性,有望作为经典诊断标准的辅助指标提高早期尘肺诊断的准确性。
[Abstract]:objective
At present, the diagnosis of pneumoconiosis mainly take the occupation history combined with imaging methods, once diagnosed, patients with pulmonary fibrosis has not reversed. Although the prevention and treatment of pneumoconiosis mainly lies in the dust concentration control of the working environment, but to explore the early diagnosis index, starting from the "two prevention", to provide scientific basis for the prevention of pneumoconiosis. Therefore, biomarkers for early diagnosis of pneumoconiosis, for early detection of suspected patients, early treatment, reduce the incidence of pneumoconiosis and the major occupation occupation rate of the tumor, which has great economic and social benefits. The theme in screening specific expression of pneumoconiosis patients serum microRNA (miRNA), to explore the serum miRNA as pneumoconiosis early biomarkers of diagnostic value.
Method
The low density TaqMan chip (TLDA) method, can be used as screening candidate serum miRNA screening diagnosis and prognosis prediction of pneumoconiosis marker, and RT-qPCR method was used for verification. Through a case-control study and statistical analysis, get clear biomarkers for screening and diagnosis of pneumoconiosis in blood, serum marker for the diagnosis of miRNA the value and receiveroperating curve to evaluate the candidate, to compare the sensitivity and specificity. The final fitting out a number of candidate miRNA for early screening, early diagnosis of pneumoconiosis, combined application of model predictive.
Result
By 1. TaqMan low density array method selected in the control group, the pneumoconiosis group between expression of 10 candidate miRNA have differences in serum by RT-qPCR method in 130 cases of pneumoconiosis stage I, II verified 22 cases of pneumoconiosis and 152 control serum samples, can be detected effectively.
2., compared with healthy control group, 6 miRNA in pneumoconiosis samples were highly expressed, which were miR-21, miR-200c, miR-16, miR-206, miR-155, miR-29a, and 1 showed low expression (miR-204), the difference was significant.
3. comparison of pneumoconiosis between stage I and II: the results showed that there was a difference in the expression of miR-204 and miR-21 in stage I and II pneumoconiosis. MiR-21 expression increased progressively in the control group and stage I and phase II pneumoconiosis, and miR-204 decreased in the control group and in stage I and II pneumoconiosis.
4. fitting 6 regression models with Logistic pneumoconiosis screening diagnosis value and good sensitivity and specificity of serum miRNA to establish the screening and diagnosis of pneumoconiosis and normal people: logitP=13.769+0.536 * miR-21 0.878 * miR-200C 0.012 * miR-16 0.111 * miR-206+0.117 * miR-155 1.192 * miR-29a, the AUC-ROC was 0.980 (95%CI:0.970-0.993, P < 0.0001).
conclusion
1. using TaqMan low density chip to draw miRNA expression profile of pneumoconiosis serum, the method is reliable and feasible.
2. serum miRNA is expected to be a marker for the diagnosis of pneumoconiosis screening, which has potential application value.
3. the progression of pneumoconiosis can be prompted by the expression level of serum miRNA.
4. combined detection of multiple serum miRNA can improve the sensitivity and specificity of pneumoconiosis diagnosis, and is expected to serve as an auxiliary index of classical diagnostic criteria to improve the accuracy of early pneumoconiosis diagnosis.

【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R135.2

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