本文選題：塵肺　切入點(diǎn)：miRNA　出處：《天津醫(yī)科大學(xué)》2013年碩士論文　論文類型：學(xué)位論文

【摘要】：目的 目前,對塵肺的診斷主要采取職業(yè)史結(jié)合影像學(xué)的方式,一旦確診,病人肺纖維化進(jìn)程已無法逆轉(zhuǎn)。雖然塵肺病的防治主要在于控制工作環(huán)境的粉塵濃度,但是探索早期的診斷指標(biāo),從“二級預(yù)防”出發(fā),為預(yù)防塵肺提供科學(xué)依據(jù)。因此,尋找塵肺早期診斷的生物標(biāo)志物,對于早期發(fā)現(xiàn)疑似病人,早治療,降低塵肺和職業(yè)性腫瘤等重大職業(yè)病的發(fā)病率,具有重大的經(jīng)濟(jì)和社會效益。本課題旨在篩選塵肺病人血清中特異表達(dá)的microRNA(miRNA),探索血清miRNA作為塵肺早期生物標(biāo)志物的診斷價值。 方法 采用TaqMan低密度芯片(TLDA)的方法,篩選出候選的可作為塵肺的篩查診斷及預(yù)后預(yù)測的血清miRNA標(biāo)志物,并采用RT-qPCR方法進(jìn)行驗(yàn)證。通過病例對照研究和統(tǒng)計分析,得到可用于塵肺篩查診斷的血清生物標(biāo)志物,并繪制受試者工作曲線評估候選的血清miRNA標(biāo)志物對塵肺的診斷價值,比較其靈敏度和特異性。最終擬合出多個候選miRNA用于塵肺早期篩查、早期診斷、預(yù)后預(yù)測的聯(lián)合應(yīng)用模型。 結(jié)果 1.通過TaqMan低密度芯片方法篩選出在正常對照組、塵肺組之間血清中有一定表達(dá)差異的10個候選miRNA,用RT-qPCR方法在Ⅰ期塵肺130例,Ⅱ期塵肺22例和對照152例血清樣本中進(jìn)行驗(yàn)證,均可檢測到有效表達(dá)。 2.與健康對照組相比,塵肺樣本中6個miRNA呈高表達(dá),分別為miR-21、 miR-200c、miR-16、miR-206、miR-155、miR-29a,1個呈低表達(dá)(miR-204),差異均有顯著性。 3. Ⅰ、Ⅱ期塵肺間的比較：結(jié)果顯示miR-204、miR-21在Ⅰ、Ⅱ期塵肺中的表達(dá)量存在差異性,miR-21在對照組以及Ⅰ、Ⅱ期塵肺中表達(dá)遞增,miR-204在對照組以及Ⅰ、Ⅱ期塵肺中表達(dá)遞減。 4.擬合6個有塵肺篩查診斷價值且靈敏度和特異性良好的血清miRNA建立塵肺和正常人群中篩查診斷的Logistic回歸模型：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為0.980(95%CI:0.970-0.993, P＜0.0001)。 結(jié)論 1.利用TaqMan低密度芯片繪制塵肺血清miRNA表達(dá)譜,方法可靠可行。 2.血清miRNA可望作為塵肺篩查診斷的標(biāo)志物,具有潛在的應(yīng)用價值。 3.通過血清miRNA的表達(dá)水平,可以提示塵肺發(fā)病進(jìn)程。 4.多個血清miRNA聯(lián)合應(yīng)用檢測可改善塵肺診斷的靈敏度和特異性,有望作為經(jīng)典診斷標(biāo)準(zhǔn)的輔助指標(biāo)提高早期塵肺診斷的準(zhǔn)確性。
[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.

【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R135.2

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