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  本文選題：鼻咽癌　切入點：DEPDC1　出處：《重慶醫(yī)科大學》2017年碩士論文　論文類型：學位論文

【摘要】：DEPDC1(DEP domain containing 1)是近年來發(fā)現(xiàn)的一個腫瘤相關基因,前期研究結(jié)果表明DEPDC1在鼻咽癌細胞周期以及癌變過程中具有重要作用。目的:為了更進一步探討其在鼻咽癌發(fā)生發(fā)展中的作用。方法:利用qRT-PCR和免疫組織化學染色分別檢測DEPDC1在33例新鮮鼻咽癌組織樣品和44例鼻咽癌組織芯片中的表達情況。人鼻咽癌細胞株CNE-1穩(wěn)定沉默DEPDC1后,流式細胞技術檢測細胞周期,CCK8檢測細胞增殖,transwell分析細胞侵襲遷移的情況,利用裸鼠成瘤試驗分析體內(nèi)成瘤能力。結(jié)果:DEPDC1在鼻咽癌組織中表達量(0.699±0.521)明顯高于鼻咽部正常組織(0.408±0.183),差異有統(tǒng)計學意義(P0.05)。DEPDC1穩(wěn)定沉默后,CCK8檢測結(jié)果表明CNE-1細胞增殖速度減慢;流式細胞檢測結(jié)果顯示,G1期細胞減少,G2/M期細胞增多,細胞周期進程受阻。劃痕及Transwell實驗結(jié)果表明,DEPDC1穩(wěn)定沉默明顯減弱了CNE-1細胞運動、侵襲及遷移能力(77.033±5.183 20.137±2.828)(107.336±5.033 26.326±1.414),差異均有統(tǒng)計學意義(P0.05)。進一步定量qRT-PCR及Western印跡檢測發(fā)現(xiàn),DEPDC1穩(wěn)定沉默導致上皮細胞標志分子Ecadherin(0.457±0.022)顯著上調(diào)、而間質(zhì)細胞標志分子Vimentin(0.780±0.063)、Ncadherin(0.780±0.063)以及EMT上游關鍵轉(zhuǎn)錄因子Twist1(0.710±0.034)下調(diào)(P0.05)。裸鼠成瘤實驗表明DEPDC1穩(wěn)定沉默后抑制了裸鼠成瘤能力,與對照組(0.23±0.03g,46.91±15.07mm3)相比,DEPDC1穩(wěn)定沉默后腫瘤重量和體積(0.56±0.17g,546.24±93.46mm3)明顯減小,差異均有統(tǒng)計學意義(P0.05)。結(jié)論:本研究成功構(gòu)建了DEPDC1穩(wěn)定沉默的鼻咽癌細胞株,與前期siRNA介導的瞬時沉默相似,DEPDC1穩(wěn)定沉默可抑制鼻咽癌細胞的增殖、侵襲遷移,并改變EMT關鍵分子的表達,為進一步探索DEPDC1在鼻咽癌中的作用機理奠定了基礎。
[Abstract]:DEPDC1(DEP domain containing 1 is a tumor-related gene discovered in recent years. Previous studies have shown that DEPDC1 plays an important role in the cell cycle and carcinogenesis of nasopharyngeal carcinoma. Objective: to further explore the role of DEPDC1 in the carcinogenesis and development of nasopharyngeal carcinoma. Methods: qRT-PCR and immunohistochemical staining were used to study the role of DEPDC1 in the development of nasopharyngeal carcinoma. The expression of DEPDC1 in 33 fresh nasopharyngeal carcinoma tissue samples and 44 nasopharyngeal carcinoma tissue microarray samples were detected respectively. Human nasopharyngeal carcinoma cell line CNE-1 was stably silenced with DEPDC1. Flow cytometric analysis of cell cycle CCK8 cell proliferation and transwell analysis of cell invasion and migration. Results the expression of w DEPDC1 in nasopharyngeal carcinoma was 0.699 鹵0.521), which was significantly higher than that in normal nasopharyngeal tissue (0.408 鹵0.183). The results showed that the proliferation rate of CNE-1 cells slowed down after stable silencing. The results of flow cytometry showed that the G 1 phase cells decreased the number of G 2 / M phase cells and the cell cycle progression was blocked. The results of scratch and Transwell showed that the stable silencing of CNE-1 cells significantly decreased the motility of CNE-1 cells. The ability of invasion and migration was 77.033 鹵5.183 20.137 鹵2.828 (107.336 鹵5.033 26.326 鹵1.414), the difference was statistically significant (P 0.05). Further quantitative qRT-PCR and Western blot analysis showed that stable silencing of DEPDC1 resulted in a marked up-regulation of Ecadherin(0.457 鹵0.022 in epithelial cells. However, Vimentin(0.780 鹵0.063 + Ncadherin (0.780 鹵0.063) and Twist1(0.710 鹵0.034 (a key transcription factor in upstream of EMT) down-regulated P0.05.The tumor-forming ability of nude mice was inhibited by DEPDC1 stable silencing, and the tumor weight and volume of DEPDC1 decreased significantly after stable silencing compared with control group (0.23 鹵0.03g / L, 46.91 鹵15.073mm), and the tumor weight and volume decreased significantly (0.56 鹵0.17g / 546.24 鹵93.46mm3), compared with that of the control group (0.23 鹵0.063 g / L, 46.91 鹵15.073mm). Conclusion: DEPDC1 stable silencing nasopharyngeal carcinoma cell line was successfully constructed. Similar to the transient silencing mediated by siRNA, DEPDC1 stable silencing can inhibit the proliferation, invasion and migration of nasopharyngeal carcinoma cells. The expression of key molecules of EMT was changed, which laid a foundation for further exploring the mechanism of DEPDC1 in nasopharyngeal carcinoma.
【學位授予單位】：重慶醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R739.63

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本文編號：1598132