本文選題：DNA甲基化水平 + 膀胱癌�。� 參考：《華僑大學(xué)》2014年博士論文

【摘要】：目的：膀胱癌是一種常見的惡性腫瘤，全球每年約15萬人死亡。迄今仍然沒有分子靶向藥物被批準(zhǔn)用于該疾病治療。TCF21（Transcription Factor21）是編碼轉(zhuǎn)錄因子21的基因，也是腫瘤抑制基因。TCF21基因甲基化在頭頸部癌、肺癌和腎癌上已有報(bào)道，然而在膀胱癌上TCF21基因甲基化表征及其臨床意義尚未得到充分的揭示，尤其是TCF21基因甲基化定量檢測(cè)在膀胱癌中未見報(bào)道。本研究探討TCF21基因甲基化與表達(dá)的相互關(guān)系，以及TCF21基因甲基化在膀胱癌中的表征，目的在于證明對(duì)膀胱癌尿脫落細(xì)胞中TCF21基因甲基化定量檢測(cè)，可以成為膀胱癌診斷和預(yù)后的重要手段之一。 方法：選擇膀胱癌細(xì)胞株5637進(jìn)行體外實(shí)驗(yàn)，用去甲基化試劑Decitabine和Zebularine處理膀胱癌細(xì)胞株5637，采用焦磷酸測(cè)序技術(shù)定量檢測(cè)膀胱癌細(xì)胞株5637中TCF21基因甲基化水平，分別采用雙標(biāo)記熒光探針技術(shù)和Western-blot方法檢測(cè)膀胱癌細(xì)胞株5637中TCF21mRNA和蛋白水平；采用焦磷酸測(cè)序技術(shù)定量檢測(cè)83例膀胱癌組織和19例正常膀胱組織中TCF21基因甲基化水平，采用Mann-Whitney U Test統(tǒng)計(jì)方法分析、比較在膀胱癌組織中TCF21基因甲基化水平在患者性別、年齡、吸煙史、腫瘤分級(jí)和分期五個(gè)臨床參數(shù)上的差異，采用Spearman Correlation統(tǒng)計(jì)方法分析在膀胱癌組織中TCF21基因甲基化水平與患者年齡、腫瘤分級(jí)和分期三個(gè)臨床參數(shù)的相關(guān)性；采用焦磷酸測(cè)序技術(shù)定量檢測(cè)33例膀胱癌尿脫落細(xì)胞和10例健康人群尿脫落細(xì)胞中TCF21基因甲基化水平，采用Mann-Whitney U Test統(tǒng)計(jì)方法分析、比較在膀胱癌尿脫落細(xì)胞中TCF21基因甲基化水平在患者性別、年齡、吸煙史、腫瘤分級(jí)和分期五個(gè)臨床參數(shù)上的差異，采用Spearman Correlation統(tǒng)計(jì)方法分析在膀胱癌尿脫落細(xì)胞中TCF21基因甲基化水平與患者年齡、腫瘤分級(jí)和分期三個(gè)臨床參數(shù)的相關(guān)性，采用受試者工作特征（ROC）曲線方法對(duì)比膀胱癌組織和膀胱癌尿脫落細(xì)胞中TCF21基因甲基化水平對(duì)診斷的潛在能力。 結(jié)果：本研究發(fā)現(xiàn)用去甲基化試劑Decitabine和Zebularine處理膀胱癌細(xì)胞株5637后，導(dǎo)致TCF21基因甲基化水平降低，TCF21mRNA和蛋白表達(dá)增加；與此同時(shí)，在膀胱癌組織中TCF21甲基化水平高于正常膀胱組織，在膀胱癌組織中TCF21甲基化水平高于腎癌組織，患者年齡對(duì)在膀胱癌組織中TCF21基因甲基化水平存在差異；在膀胱癌尿脫落細(xì)胞中TCF21甲基化水平高于健康人群尿脫落細(xì)胞，患者腫瘤分級(jí)對(duì)在膀胱癌尿脫落細(xì)胞中TCF21基因甲基化水平存在差異，患者腫瘤分級(jí)與在膀胱癌尿脫落細(xì)胞中TCF21基因甲基化水平相關(guān)，在膀胱癌組織和膀胱癌尿脫落細(xì)胞中TCF21的ROC曲線下面積均高于參考值，但在膀胱癌尿脫落細(xì)胞中TCF21特異性高于膀胱癌組織。 結(jié)論：本研究證明TCF21基因CpG島的異常甲基化在調(diào)節(jié)TCF21基因的表達(dá)中起重要作用；TCF21基因可以作為是膀胱癌診斷和預(yù)后的檢測(cè)的標(biāo)志物；對(duì)膀胱癌尿脫落細(xì)胞中TCF21基因甲基化定量檢測(cè)，有可能成為膀胱癌診斷和預(yù)后的重要手段之一。
[Abstract]:Objective: bladder cancer is a common malignant tumor with about 150 thousand people worldwide each year. There are still no molecular targeting drugs approved for the treatment of.TCF21 (Transcription Factor21), a gene encoding transcription factor 21, and a tumor suppressor gene.TCF21 methylation in the head and neck cancer, lung cancer and kidney cancer. However, the methylation of TCF21 gene and its clinical significance in bladder cancer have not been fully revealed, especially the quantitative methylation detection of TCF21 gene has not been reported in bladder cancer. The relationship between the methylation and expression of the TCF21 gene and the characterization of the methylation of the TCF21 gene in bladder cancer are discussed in this study. Quantitative detection of TCF21 gene methylation in cystadenocarcinoma exfoliated cells can be one of the important means for diagnosis and prognosis of bladder cancer.
Methods: bladder cancer cell line 5637 was selected in vitro, and bladder cancer cell line 5637 was treated with demethylation reagent Decitabine and Zebularine. The methylation level of TCF21 gene in bladder cancer cell line 5637 was detected by pyrosequencing technology. The double labeling fluorescence probe technique and Western-blot method were used to detect bladder cancer. The level of TCF21mRNA and protein in cell line 5637 and the methylation level of TCF21 gene in 83 bladder cancer tissues and 19 normal bladder tissues were quantified by pyrosequencing technology. The Mann-Whitney U Test statistical method was used to compare the TCF21 gene methylation water in the bladder cancer tissue for the patient's sex, age, smoking history and tumor classification. The Spearman Correlation statistical method was used to analyze the correlation between the level of TCF21 gene methylation of the TCF21 gene in the bladder cancer tissues and the three clinical parameters of the tumor classification and staging, and the quantitative detection of urine exfoliation in 33 cases of bladder cancer and 10 healthy people by pyrosequencing. The methylation level of TCF21 gene in the cells was analyzed by Mann-Whitney U Test statistical method, and the difference in the five clinical parameters of the level of methylation of TCF21 gene in the urinary exfoliated cells of bladder cancer was compared with the sex, age, smoking history, tumor classification and staging, and the urine exfoliative cells of bladder cancer were analyzed by Spearman Correlation. The correlation between the level of methylation of the TCF21 gene and the three clinical parameters of the patient's age, tumor classification and staging, and the potential ability of the TCF21 gene methylation in bladder cancer and urinary exfoliative cells to compare the diagnostic potential of the bladder cancer tissue and urinary bladder cancer cells were compared by the method of the subject work feature (ROC) curve.
Results: after treating bladder cancer cell line 5637 with demethylation reagents Decitabine and Zebularine, the methylation level of TCF21 gene was reduced and the expression of TCF21mRNA and protein increased. At the same time, the level of TCF21 methylation in bladder cancer tissues was higher than that of normal bladder tissue, and the level of TCF21 methylation in bladder cancer tissues was higher than that in bladder cancer tissue. The level of methylation of TCF21 gene in bladder cancer tissues is different in the patients with renal carcinoma. The level of TCF21 methylation in urinary exfoliated cells of bladder cancer is higher than that of the healthy people. The level of TCF21 gene methylation in the urinary exfoliative cells of bladder cancer is different in patients with bladder cancer, and the tumor classification and bladder cancer in the patients with bladder cancer are different. The level of TCF21 gene methylation in urine exfoliated cells was higher than that of the reference value in the ROC curve of TCF21 in bladder cancer tissue and bladder cancer urine exfoliative cells, but the specificity of TCF21 in bladder cancer urine exfoliated cells was higher than that of bladder cancer tissue.
Conclusion: This study proves that the abnormal methylation of the TCF21 gene CpG Island plays an important role in the regulation of the expression of TCF21 gene, and the TCF21 gene can be used as a marker for the diagnosis and prognosis of bladder cancer, and the quantitative detection of TCF21 gene in the urine exfoliated cells of bladder cancer can be an important hand for the diagnosis and prognosis of bladder cancer. One of the segments.

【學(xué)位授予單位】：華僑大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類號(hào)】：R737.14

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本文編號(hào)：1787801