【摘要】：目的：基于高通量測(cè)序技術(shù)、RT-qPCR和免疫組化技術(shù),從轉(zhuǎn)錄層面和蛋白表達(dá)角度探索膀胱尿路上皮癌發(fā)生發(fā)展的可能機(jī)制,篩選出膀胱腫瘤早期診斷和監(jiān)測(cè)的分子標(biāo)記。 方法：通過(guò)高通量測(cè)序技術(shù)對(duì)一膀胱尿路上皮癌的患者進(jìn)行轉(zhuǎn)錄組測(cè)序和生物信息學(xué)分析,篩選到候選的差異表達(dá)分子,同時(shí)收集31對(duì)膀胱尿路上皮癌腫瘤組織和癌旁非腫瘤組織樣品,采用RT-qPCR驗(yàn)證這些差異表達(dá)分子CDH1、VEGFA、PTPRF和CLDN7的表達(dá)水平。同時(shí)對(duì)CDH1和CLDN7進(jìn)行免疫組化進(jìn)行檢測(cè),探討這些分子在腫瘤的發(fā)生發(fā)展中的分子機(jī)制和臨床意義。 結(jié)果：1.CDH1、VEGFA、PTPRF和CLDN7可能成為候選膀胱癌早期診斷和監(jiān)測(cè)的分子標(biāo)記；CDH1和CLDN7mRNA在腫瘤組織相對(duì)癌旁非腫瘤組織表達(dá)上調(diào),但差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)；2.VEGFA在腫瘤組織中相對(duì)癌旁非腫瘤組織表達(dá)上調(diào),差異有統(tǒng)計(jì)學(xué)意義(P0.05);3. CDH1、 VEGFA、PTPRF和CLDN7分別與腫瘤復(fù)發(fā)關(guān)系作多元Logistic回歸分析,結(jié)果發(fā)現(xiàn)PTPRF表達(dá)作為腫瘤復(fù)發(fā)因素進(jìn)入模型(P=0.002),說(shuō)明PTPRF和腫瘤復(fù)發(fā)關(guān)系密切；4.CDH1和CLDN7免疫組化顯示：CDH1在膀胱腫瘤組織和癌旁非腫瘤組織蛋白表達(dá)中未發(fā)現(xiàn)有明顯統(tǒng)計(jì)學(xué)意義差異(P0.05),膀胱尿路上皮癌腫瘤組織CLDN7表達(dá)上調(diào),具有統(tǒng)計(jì)學(xué)意義(P0.05)。 結(jié)論：1.CLDN7、VEGFA和PTPRF等差異表達(dá)基因被納入可能的膀胱癌分子早期診斷和治療的分子標(biāo)記候選集；2.VEGFA可能作為候選的膀胱腫瘤診斷的分子標(biāo)記,PTPRF可能與膀胱腫瘤的復(fù)發(fā)關(guān)系比較密切。
[Abstract]:Objective: to explore the possible mechanism of bladder urothelial carcinoma from transcriptional level and protein expression based on RT-qPCR and immunohistochemistry, and to screen out the molecular markers for early diagnosis and monitoring of bladder cancer. Methods: transcriptome sequencing and bioinformatics analysis were carried out in a patient with bladder urothelial carcinoma by high-throughput sequencing technique, and candidate differentially expressed molecules were screened. At the same time, 31 samples of bladder urothelial carcinoma were collected, and the expression levels of PTPRF and CLDN7 of CDH1VEGFA-PTPRF and CLDN7 were verified by RT-qPCR. At the same time, CDH1 and CLDN7 were detected by immunohistochemistry to explore the molecular mechanism and clinical significance of these molecules in tumorigenesis and development. Results: 1. The expression of CDH1 and CLDN7 might be the molecular markers for early diagnosis and monitoring of bladder cancer, but the expression of CDH1 and CLDN7mRNA in tumor tissues was significantly higher than that in non-tumor tissues adjacent to tumor tissues, but the difference was not statistically significant (P0.05). The expression of 2.VEGFA was up-regulated in tumor tissues relative to non-tumor tissues adjacent to cancer, and the difference was statistically significant (P0.05). The multivariate Logistic regression analysis showed that the expression of PTPRF as a tumor recurrence factor entered the model (P0. 002), indicating the close relationship between PTPRF and tumor recurrence. 4.CDH1 and CLDN7 immunohistochemical staining showed that there was no significant difference in the expression of CLDN7 between bladder tumor tissue and adjacent non-tumor tissue (P0.05), but the expression of CLDN7 was up-regulated in bladder urothelial carcinoma tissue (P0.05). Conclusion: 1. The differential expression genes such as VEGFA and PTPRF were included in the molecular marker candidate for early diagnosis and treatment of bladder cancer 2.VEGFA may be used as a molecular marker for the diagnosis of bladder cancer, which may be closely related to the recurrence of bladder cancer.
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R737.14

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