miR-613在食管鳞癌诊断和预后中的作用

发布时间：2018-06-15 21:52

本文选题：食管鳞状细胞癌 + miR-613　；参考：《山东大学》2017年博士论文

【摘要】：目的:食管癌是最常见的消化道恶性肿瘤之一,在未来10年发生率会不断增高。患者的预后较差,五年生存率仅为150%-250%左右。对于食管癌标志物的探索是近几年迅速发展的研究领域,对于食管癌的早期诊断以及预后评估有重要作用。近期大量研究表明,血小板计数、肿瘤长度以及营养状态等指标与食管癌患者的预后密切相关。食管癌患者的早期诊断对于改善患者预后意义重大。miRNA的表达水平高低可以作为肿瘤的诊断和预后指标,用于评价肿瘤发生、进展以及治疗效果。例如,血清miR-25、miR-223和miR-375可以作为食管鳞癌的诊断指标。血清miR-24和miR-200c同样可以作为评价患者化疗和同步放化疗疗效的无创性指标。miR-613在不同的肿瘤中发挥不同的作用,在乳头状甲状腺癌中低表达,在胃癌中却为高表达。可以靶向调节FNA基因以及Wnt通路。然而,miR-613在食管癌中的表达水平及其作用机制尚不明确。本文目的:探讨miR-613在食管癌肿瘤组织以及血清中的表达水平,并进一步探讨了其临床诊断和预后评价价值。资料和方法:我们纳入了齐鲁医院2014-10到2015-03之间胸外科75名拟接受手术治疗的食管癌患者的术前血清样本,同时收集了年龄及性别与之配对的正常人群的血清样本作为对照组。同时,我们收集了其中60位患者的食管癌组织及其周围正常组织(15例患者未行手术治疗,因拒绝手术或身体状态不适宜手术),术后30min内收集好肿瘤组织和癌旁组织,用液氮转移至-80度冰箱中保存。我们还收集了 2009年齐鲁医院病理科的109名食管鳞癌患者的福尔马林固定石蜡包埋的组织标本(FFPE)。所有患者均经手术或食管镜活检诊断为食管鳞癌。患者均未行术前抗肿瘤治疗。我们统计分析了患者的年龄、性别、吸烟及饮酒情况、肿瘤长度、位置、浸润深度(T分期)、淋巴结转移程度(N分期)、远处转移(M分期)以及分化程度总RNA提取及qRT-PCR检测RT反应体系总共25ul,qPCR体系为20ul,RNU6B和miR-16分别作为组织和血清中的内参。组织和血清中miR-613的相对表达量采用2-△ △Ct计算,FFPE中的表达量采用2-△ Ct计算。实验结果:1、食管鳞癌患者组织中miR-613的表达情况我们采用qRT-PCR方法分析肿瘤组织和癌旁正常组织中miR-613表达量。与正常组织相比,肿瘤组织miR-613的表达呈下调趋势(0.34±0.56vs.1.08±0.07,P0.001)。同时分析miR-613的表达水平与患者临床特征之间的关系,我们发现miR-613的表达水平与患者T分期呈负相关(P=0.008)。2、血清miR-613的表达水平及其诊断价值我们首先假设组织miR-613的表达水平可能会影响血清中的表达情况。实验结果显示患者血清miR-613也为低表达趋势(0.89±0.73 vs.1.71 士 1.03,P0.001)。血清miR-613的表达水平与患者N分期呈明显负相关(P=0.038),而与其他指标没有明显相关性。我们分析了患者食管癌组织miR-613与血清miR-613表达的相关性,Pearson相关系数为0.812(P0.001)。我们分析了血清miR-613的诊断价值,曲线下面积(AUC)为0.767±0.040。Yuden指数,即cut-off值为0.875,R0C曲线的敏感性和特异性值分别为81.3%and 62.7%。对早期患者数据的进一步分析发现,AUC值为0.728±0.052(P0.001)。3、miR-613在食管鳞癌患者预后的预测价值我们收集了 109名食管癌患者的FFPE标本,miR-613的平均表达水平为0.0016(cut-off值)。miR-613的表达水平与患者临床特征之间没有明显相关性。患者的平均生存时间为39个月(分布区间为6-81月)。截至末次随访,35名患者(32.11%)仍健在,而74名患者(67.89%)已去世。K-M曲线分析提示miR-613低表达组患者的五年总生存时间(0S)及无进展生存时间(PFS)明显较短(P= 0.018和P=0.035,respectively,)。进一步的多因素分析提示miR-613是患者 OS(HR(95%CI)0.591(0.336-0.954),P=0.031)和 PFS(HR(95%CI)0.656(0.485-0.888),P = 0.006,)良好的预后指标。结论:(1)与正常组织相比,肿瘤组织miR-613的表达呈下调趋势;(2)肿瘤组织中miR-613的表达水平与患者T分期呈负相关;(3)患者食管癌组织miR-613与血清miR-613表达的一定的正相关性,血清miR-613的表达水平与患者N分期呈明显负相关,并且具有一定的诊断价值;(4)miR-613低表达提示患者预后较差。
[Abstract]:Objective: esophageal cancer is one of the most common digestive tract malignant tumors. The incidence of esophageal cancer in the next 10 years is increasing. The prognosis of the patients is poor and the five year survival rate is only about 150%-250%. The exploration of the esophageal cancer markers is a rapid development in recent years. It has an important role in the early diagnosis and prognosis evaluation of esophageal cancer. A large number of studies have shown that the platelet count, tumor length, and nutritional status are closely related to the prognosis of patients with esophageal cancer. Early diagnosis of patients with esophageal cancer can be used as a diagnostic and prognostic marker for the improvement of the prognosis of patients with.MiRNA, which is used to evaluate the occurrence, progress and therapeutic effect of cancer. For example, serum miR-25, miR-223, and miR-375 can be used as diagnostic indicators for squamous cell carcinoma of the esophagus. Serum miR-24 and miR-200c can also be used as a noninvasive index to evaluate the efficacy of chemotherapy and concurrent chemoradiotherapy..miR-613 plays a different role in different tumors, low expression in papillary thyroid carcinoma and high expression in gastric cancer. It can be targeted to regulate FNA gene and Wnt pathway. However, the expression level and mechanism of miR-613 in esophageal cancer are not clear. Objective: To explore the expression level of miR-613 in the tumor tissues and serum of esophageal cancer, and to further explore the value of its clinical diagnosis and prognosis evaluation. The preoperative serum samples of 75 patients with surgical treatment of esophageal cancer were collected from 2014-10 to 2015-03 in the hospital, and serum samples of age and sex matched normal people were collected as the control group. At the same time, we collected 60 of the patients' esophageal cancer tissues and its surrounding normal tissues (15 patients were not treated by surgery. " The tumor tissue and paracancerous tissue were collected and transferred to the -80 degree refrigerator at 30min after the operation. We also collected the formalin fixed paraffin embedded tissue specimens (FFPE) of 109 patients with esophageal squamous cell carcinoma in Qilu Hospital in 2009. All patients were operated by surgery or Esophageal squamous cell carcinoma was diagnosed as squamous cell carcinoma of the esophagus. All patients were not treated with preoperative antitumor treatment. We statistically analyzed age, sex, smoking and drinking, tumor length, location, depth of infiltration (T staging), lymph node metastasis (N staging), distant metastasis (M phase), total RNA extraction and qRT-PCR detection of RT reaction system. A total of 25ul, qPCR system was 20ul, RNU6B and miR-16 were used as internal parameters in tissue and serum respectively. The relative expression of miR-613 in tissue and serum was calculated by 2- Delta Ct, and the expression in FFPE was calculated by 2- Delta Ct. Experimental results: 1, the expression of miR-613 in tissues of patients with esophageal squamous cell carcinoma was analyzed by the method of analysis of tumor tissue and Compared with normal tissue, the expression of miR-613 in the tumor tissue was down downward (0.34 + 0.56vs.1.08 + 0.07, P0.001). Meanwhile, the relationship between the expression level of miR-613 and the clinical features of the patients was also analyzed. We found that the expression level of miR-613 was negatively correlated with the T stage of the patients (P=0.008).2 and serum miR-613. We first hypothesized that the expression level of the tissue miR-613 might affect the expression in the serum. The experimental results showed that the serum miR-613 was also a low expression trend (0.89 + 0.73 vs.1.71 1.03, P0.001). The expression level of serum miR-613 was negatively correlated with the patients' N staging (P=0.038), but with the other fingers. The correlation between miR-613 and serum miR-613 expression was analyzed. The Pearson correlation coefficient was 0.812 (P0.001). We analyzed the diagnostic value of serum miR-613. The area under the curve (AUC) was 0.767 + 0.040.Yuden index, that is, cut-off value was 0.875, and the sensitivity and specificity of R0C curve were 8, respectively. 1.3%and 62.7%.'s further analysis of early patient data found that the AUC value was 0.728 + 0.052 (P0.001).3. The predictive value of miR-613 in the prognosis of esophageal squamous cell carcinoma we collected the FFPE specimens of 109 esophageal cancer patients. The average expression level of miR-613 was 0.0016 (cut-off).MiR-613 expression and the patient's clinical characteristics were not between them. The average survival time of the patients was 39 months (6-81 months in the distribution interval). 35 patients (32.11%) were still alive at the end of the last follow-up, and 74 patients (67.89%) had passed the.K-M curve analysis. The total survival time (0S) and the progression free survival time (PFS) in the miR-613 low expression group were significantly shorter (P= 0.018 and P=0.035, respe, respe). Ctively. Further multivariate analysis suggested that miR-613 was a good prognostic indicator for patients with OS (HR (95%CI) 0.591 (0.336-0.954) 0.591 (0.336-0.954), P=0.031) and PFS (HR (95%CI) 0.656 (0.485-0.888), P = 0.006). Conclusion: (1) the expression of tumor tissue was down downward trend compared with normal tissue; (2) the expression level of the tumor tissue was compared with the patient. There was a negative correlation. (3) there was a positive correlation between the miR-613 and the expression of serum miR-613 in the patients with esophageal cancer. The expression level of serum miR-613 was negatively correlated with the N staging of the patients, and was of certain diagnostic value; (4) the low expression of miR-613 suggested that the prognosis of the patients was poor.
【学位授予单位】：山东大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R735.1

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