本文選題：食管鱗狀細(xì)胞癌 + miR-613　； 參考：《山東大學(xué)》2017年博士論文

【摘要】：目的:食管癌是最常見(jiàn)的消化道惡性腫瘤之一,在未來(lái)10年發(fā)生率會(huì)不斷增高。患者的預(yù)后較差,五年生存率僅為150%-250%左右。對(duì)于食管癌標(biāo)志物的探索是近幾年迅速發(fā)展的研究領(lǐng)域,對(duì)于食管癌的早期診斷以及預(yù)后評(píng)估有重要作用。近期大量研究表明,血小板計(jì)數(shù)、腫瘤長(zhǎng)度以及營(yíng)養(yǎng)狀態(tài)等指標(biāo)與食管癌患者的預(yù)后密切相關(guān)。食管癌患者的早期診斷對(duì)于改善患者預(yù)后意義重大。miRNA的表達(dá)水平高低可以作為腫瘤的診斷和預(yù)后指標(biāo),用于評(píng)價(jià)腫瘤發(fā)生、進(jìn)展以及治療效果。例如,血清miR-25、miR-223和miR-375可以作為食管鱗癌的診斷指標(biāo)。血清miR-24和miR-200c同樣可以作為評(píng)價(jià)患者化療和同步放化療療效的無(wú)創(chuàng)性指標(biāo)。miR-613在不同的腫瘤中發(fā)揮不同的作用,在乳頭狀甲狀腺癌中低表達(dá),在胃癌中卻為高表達(dá)�？梢园邢蛘{(diào)節(jié)FNA基因以及Wnt通路。然而,miR-613在食管癌中的表達(dá)水平及其作用機(jī)制尚不明確。本文目的:探討miR-613在食管癌腫瘤組織以及血清中的表達(dá)水平,并進(jìn)一步探討了其臨床診斷和預(yù)后評(píng)價(jià)價(jià)值。資料和方法:我們納入了齊魯醫(yī)院2014-10到2015-03之間胸外科75名擬接受手術(shù)治療的食管癌患者的術(shù)前血清樣本,同時(shí)收集了年齡及性別與之配對(duì)的正常人群的血清樣本作為對(duì)照組。同時(shí),我們收集了其中60位患者的食管癌組織及其周?chē)＝M織(15例患者未行手術(shù)治療,因拒絕手術(shù)或身體狀態(tài)不適宜手術(shù)),術(shù)后30min內(nèi)收集好腫瘤組織和癌旁組織,用液氮轉(zhuǎn)移至-80度冰箱中保存。我們還收集了 2009年齊魯醫(yī)院病理科的109名食管鱗癌患者的福爾馬林固定石蠟包埋的組織標(biāo)本(FFPE)。所有患者均經(jīng)手術(shù)或食管鏡活檢診斷為食管鱗癌�；颊呔葱行g(shù)前抗腫瘤治療。我們統(tǒng)計(jì)分析了患者的年齡、性別、吸煙及飲酒情況、腫瘤長(zhǎng)度、位置、浸潤(rùn)深度(T分期)、淋巴結(jié)轉(zhuǎn)移程度(N分期)、遠(yuǎn)處轉(zhuǎn)移(M分期)以及分化程度總RNA提取及qRT-PCR檢測(cè)RT反應(yīng)體系總共25ul,qPCR體系為20ul,RNU6B和miR-16分別作為組織和血清中的內(nèi)參。組織和血清中miR-613的相對(duì)表達(dá)量采用2-△ △Ct計(jì)算,FFPE中的表達(dá)量采用2-△ Ct計(jì)算。實(shí)驗(yàn)結(jié)果:1、食管鱗癌患者組織中miR-613的表達(dá)情況我們采用qRT-PCR方法分析腫瘤組織和癌旁正常組織中miR-613表達(dá)量。與正常組織相比,腫瘤組織miR-613的表達(dá)呈下調(diào)趨勢(shì)(0.34±0.56vs.1.08±0.07,P0.001)。同時(shí)分析miR-613的表達(dá)水平與患者臨床特征之間的關(guān)系,我們發(fā)現(xiàn)miR-613的表達(dá)水平與患者T分期呈負(fù)相關(guān)(P=0.008)。2、血清miR-613的表達(dá)水平及其診斷價(jià)值我們首先假設(shè)組織miR-613的表達(dá)水平可能會(huì)影響血清中的表達(dá)情況。實(shí)驗(yàn)結(jié)果顯示患者血清miR-613也為低表達(dá)趨勢(shì)(0.89±0.73 vs.1.71 士 1.03,P0.001)。血清miR-613的表達(dá)水平與患者N分期呈明顯負(fù)相關(guān)(P=0.038),而與其他指標(biāo)沒(méi)有明顯相關(guān)性。我們分析了患者食管癌組織miR-613與血清miR-613表達(dá)的相關(guān)性,Pearson相關(guān)系數(shù)為0.812(P0.001)。我們分析了血清miR-613的診斷價(jià)值,曲線下面積(AUC)為0.767±0.040。Yuden指數(shù),即cut-off值為0.875,R0C曲線的敏感性和特異性值分別為81.3%and 62.7%。對(duì)早期患者數(shù)據(jù)的進(jìn)一步分析發(fā)現(xiàn),AUC值為0.728±0.052(P0.001)。3、miR-613在食管鱗癌患者預(yù)后的預(yù)測(cè)價(jià)值我們收集了 109名食管癌患者的FFPE標(biāo)本,miR-613的平均表達(dá)水平為0.0016(cut-off值)。miR-613的表達(dá)水平與患者臨床特征之間沒(méi)有明顯相關(guān)性�；颊叩钠骄鏁r(shí)間為39個(gè)月(分布區(qū)間為6-81月)。截至末次隨訪,35名患者(32.11%)仍健在,而74名患者(67.89%)已去世。K-M曲線分析提示miR-613低表達(dá)組患者的五年總生存時(shí)間(0S)及無(wú)進(jìn)展生存時(shí)間(PFS)明顯較短(P= 0.018和P=0.035,respectively,)。進(jìn)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,)良好的預(yù)后指標(biāo)。結(jié)論:(1)與正常組織相比,腫瘤組織miR-613的表達(dá)呈下調(diào)趨勢(shì);(2)腫瘤組織中miR-613的表達(dá)水平與患者T分期呈負(fù)相關(guān);(3)患者食管癌組織miR-613與血清miR-613表達(dá)的一定的正相關(guān)性,血清miR-613的表達(dá)水平與患者N分期呈明顯負(fù)相關(guān),并且具有一定的診斷價(jià)值;(4)miR-613低表達(dá)提示患者預(yù)后較差。
[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.
【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R735.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 殷言言;王立東;趙學(xué)科;宋昕;黃佳;韓雪娜;馮常煒;;早期食管癌患者臨床癥狀與臨床病理特征及生存狀況的關(guān)系分析[J];河南醫(yī)學(xué)研究;2015年05期

2 陳曉林;歐陽(yáng)小平;黃玉民;陶玉堅(jiān);;MicroRNA-182在非小細(xì)胞肺癌中的表達(dá)及臨床意義[J];臨床肺科雜志;2015年06期

3 Ming Li;Xiao-Yan He;Zhi-Mei Zhang;Shuo Li;Li-Hua Ren;Ri-Sheng Cao;Ya-Dong Feng;Yin-Lin Ji;Ye Zhao;Rui-Hua Shi;;Micro RNA-1290 promotes esophageal squamous cell carcinoma cell proliferation and metastasis[J];World Journal of Gastroenterology;2015年11期

4 Kyle J Napier;Mary Scheerer;Subhasis Misra;;Esophageal cancer: A Review of epidemiology, pathogenesis, staging workup and treatment modalities[J];World Journal of Gastrointestinal Oncology;2014年05期

5 滿朝來(lái);楊美玲;;體液循環(huán)microRNA的研究進(jìn)展[J];中國(guó)生物工程雜志;2014年02期

6 王江峰;凌志強(qiáng);毛偉敏;;MicroRNA-29b在食管鱗狀細(xì)胞癌中的表達(dá)及臨床意義[J];實(shí)用腫瘤雜志;2013年05期

7 羅君;凌志強(qiáng);彭兵鋒;袁嘉敏;鄭智國(guó);毛偉敏;;MicroRNA-31在食管鱗狀細(xì)胞癌中的表達(dá)及其與預(yù)后的關(guān)系[J];中國(guó)癌癥雜志;2013年07期

8 王確;周儒白;田李星;盧鈾;;血管內(nèi)皮生長(zhǎng)因子與食管癌預(yù)后相關(guān)性的Meta分析[J];中國(guó)循證醫(yī)學(xué)雜志;2013年05期

9 彭學(xué)宏;陳暖;李克;鄭邦希;;食管癌外周血miRNA的研究[J];中國(guó)校醫(yī);2013年05期

10 Peng Ren;Zhen-Tao Yu;Li Xiu;Mei Wang;Hua-Min Liu;;Elevated serum levels of human relaxin-2 in patients with esophageal squamous cell carcinoma[J];World Journal of Gastroenterology;2013年15期

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本文編號(hào)：2023771