本文選題：食管腺癌 + Ouabain��； 參考：《福建醫(yī)科大學(xué)》2016年碩士論文

【摘要】：目的研究Ouabain對于食管腺癌細(xì)胞的生長抑制作用,探索其作用的基因靶點MKK6及其下游基因SOX9對食管腺癌細(xì)胞體外生長及體內(nèi)成瘤狀況的影響。方法1考察1043種藥物對食管腺癌細(xì)胞體外生長的抑制情況,獲得候選藥物Ouabain,并利用腫瘤細(xì)胞體外生長曲線和體內(nèi)成瘤實驗來驗證該藥物對的食管腺癌細(xì)胞生長的干擾作用。2通過RNA測序技術(shù)分析經(jīng)Ouabain作用過的食管腺癌細(xì)胞內(nèi)基因表達量的變化,獲得潛在靶標(biāo)基因MKK6,并運用實時熒光定量PCR和Western Blot檢驗該基因在Ouabain作用的食管腺癌細(xì)胞及腫瘤中的表達情況。3應(yīng)用慢病毒在食管腺癌細(xì)胞導(dǎo)入可介導(dǎo)MKK6基因敲落的sh RNA,并利用實時熒光定量PCR和Western Blot檢驗該基因在腫瘤細(xì)胞中的敲落效率。進一步分析MKK6基因敲除對食管腺癌細(xì)胞體外生長狀況的影響,以及小鼠體內(nèi)成瘤情況的作用。4基于相關(guān)信號通路理論,利用實時熒光定量PCR甄選出受MKK6調(diào)控的下游靶標(biāo)基因SOX9,進而驗證Ouabain作用及MKK6敲落對SOX9基因表達水平及蛋白翻譯水平的影響。5利用CRISPR-Cas9技術(shù)于食管腺癌細(xì)胞內(nèi)敲除SOX9,通過對比空白對照組與SOX9基因敲除組在腫瘤細(xì)胞體外生長曲線和體內(nèi)成瘤實驗中的差別,探求SOX9在食管腺癌細(xì)胞增殖中發(fā)揮的作用。結(jié)果1 Ouabain對食管腺癌細(xì)胞的抑制作用:藥物篩選獲得的候選藥物Ouabain,可抑制食管腺癌細(xì)胞OE33及OE19的體外增殖情況,腹腔注射給藥可限制NSC小鼠體內(nèi)成瘤的重量及體積大小。2 Ouabain對MKK6基因轉(zhuǎn)錄與翻譯水平的影響:RNA測序顯示經(jīng)Ouabain作用的OE33細(xì)胞內(nèi)MKK6基因表達水平下降明顯,蛋白翻譯水平相較空白對照組也有所下降,OE19細(xì)胞內(nèi)影響亦同。3體外敲落MKK6基因?qū)κ彻芟侔┘?xì)胞的抑制作用:sh RNA介導(dǎo)的食管腺癌細(xì)胞內(nèi)MKK6基因敲落,可抑制食管腺癌細(xì)胞OE33及OE19的體外增殖情況,減小細(xì)胞在裸鼠體內(nèi)的成瘤的重量及體積大小。4食管腺癌細(xì)胞內(nèi)MKK6對SOX9的調(diào)控作用:基因敲落MKK6可降低OE33及0E19細(xì)胞內(nèi)轉(zhuǎn)錄因子SOX9的基因表達水平,和蛋白質(zhì)翻譯水平;5 SOX9對于食管腺癌細(xì)胞增殖的影響:食管腺癌細(xì)胞OE19內(nèi)敲除SOX9基因,可抑制其在體外的增殖和體內(nèi)的成瘤。結(jié)論1 Ouabain可抑制食管腺癌細(xì)胞體外增殖及體內(nèi)成瘤;2 Ouabain可降低食管腺癌細(xì)胞及小鼠腫瘤內(nèi)MKK6基因表達與蛋白翻譯水平;3基因敲落MKK6可抑制食管腺癌細(xì)胞體外增殖及體內(nèi)成瘤;4基因敲落MKK6可降低下游基因SOX9的基因表達與蛋白翻譯水平;5基因敲除SOX9可影響食管腺癌細(xì)胞體外增殖及體內(nèi)成瘤。
[Abstract]:Objective to study the inhibitory effect of Ouabain on the growth of esophageal adenocarcinoma cells, and to explore the effect of MKK6 and its downstream gene SOX9 on the growth of esophageal adenocarcinoma cells in vitro and tumor formation in vivo. Methods 1 the inhibition of 1043 drugs on the growth of esophageal adenocarcinoma cells in vitro was investigated. A candidate drug Ouabain was obtained, and the tumor cell growth curve in vitro and in vivo tumorigenesis test were used to verify the interference effect of the drug on the growth of esophageal adenocarcinoma cells. 2. Analysis of Ouabain treated esophageal adenocarcinoma cells by RNA sequencing technique Changes in the amount of internal gene expression, The potential target gene MKK6 was obtained, and the expression of MKK6 gene in Ouabain treated esophageal adenocarcinoma cells and tumors was detected by real-time fluorescent quantitative PCR and Western blot. 3. 3 Lentivirus transfection into esophageal adenocarcinoma cells can mediate MKK6 gene knockout. The knockout efficiency of the gene in tumor cells was detected by real-time fluorescent quantitative PCR and Western Blot. To further analyze the effect of MKK6 knockout on the growth of esophageal adenocarcinoma cells in vitro, and the role of tumorigenesis in mice based on the theory of related signal pathway. The downstream target gene SOX9 regulated by MKK6 was selected by real-time fluorescent quantitative PCR, and the effect of Ouabain and MKK6 knockout on SOX9 gene expression and protein translation was verified. 5 the CRISPR-Cas9 technique was used to knockout SOX9 in esophageal adenocarcinoma cells. By comparing the difference between blank control group and SOX9 gene knockout group in tumor cell growth curve in vitro and in vivo tumorigenesis test, To explore the role of SOX 9 in the proliferation of esophageal adenocarcinoma cells. Results 1 the inhibitory effect of Ouabain on esophageal adenocarcinoma cells: Ouabain, a candidate drug obtained by drug screening, could inhibit the proliferation of esophageal adenocarcinoma cells OE33 and OE19 in vitro. The effect of intraperitoneal injection on the transcription and translation of MKK6 gene in NSC mice by limiting the weight and volume of tumor. 2. The expression of MKK6 gene in OE33 cells induced by Ouabain was significantly decreased. The effect of MKK6 gene knockout on esophageal adenocarcinoma cells was also similar to that of the control group. The inhibitory effect of MKK6 gene knockout on esophageal adenocarcinoma cells was also similar to that of the control group. The effect of MKK6 gene knockout on esophageal adenocarcinoma cells mediated by 20% sh RNA was also similar to that of MKK6 gene knockout in vitro. Can inhibit the proliferation of esophageal adenocarcinoma cells OE33 and OE19 in vitro, Decrease the weight and volume of tumor in nude mice. 4 the regulation of SOX9 by MKK6 in esophageal adenocarcinoma cells: knockout MKK6 can decrease the expression of SOX9 in OE33 and 0E19 cells. Effects of 5SOX9 on the proliferation of esophageal adenocarcinoma cells: knockout of SOX9 gene in esophageal adenocarcinoma OE19 can inhibit its proliferation in vitro and tumorigenesis in vivo. Conclusion 1 Ouabain can inhibit the proliferation of esophageal adenocarcinoma cells in vitro and tumorigenesis in vivo. 2. Ouabain can reduce the expression of MKK6 gene and the level of protein translation in esophageal adenocarcinoma cells and mouse tumors. The knockout of MKK6 gene can inhibit the proliferation of esophageal adenocarcinoma cells in vitro. The expression of SOX9 gene and protein translation of downstream gene SOX9 can be reduced by knockout MKK6 of tumor-forming 4 gene in vivo. The knockout of SOX9 gene may affect the proliferation of esophageal adenocarcinoma cells in vitro and tumorigenesis in vivo.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：R735.1

【相似文獻】

相關(guān)期刊論文 前10條

1 李易,韓盛璽,沈通良;食管腺癌41例分析[J];四川醫(yī)學(xué);2002年06期

2 章宏,厲有名,王偉;食管腺癌臨床流行病學(xué)研究[J];中華消化雜志;2003年11期

3 Lepage C.;Bouvier A.-M.;Manfredi S. ;J. Faivre;王曉君;;特定人群食管腺癌的發(fā)病趨勢與治療[J];世界核心醫(yī)學(xué)期刊文摘(胃腸病學(xué)分冊);2006年Z1期

4 王純巍;反流增加食管腺癌的危險[J];英國醫(yī)學(xué)雜志(中文版);1999年04期

5 黎榮光,鄒浩元,黃國棟,張漢雄;食管腺癌放射治療20例報告[J];腫瘤研究與臨床;2001年04期

6 文載律,羅偉,李文才,梁麗麗,季磊;食管腺癌77例臨床病理分析[J];診斷病理學(xué)雜志;2002年03期

7 莫宏波,劉譽,陳萬群;食管腺癌生物標(biāo)記物的研究進展[J];腫瘤防治研究;2005年08期

8 孫永剛;周鋼;王偉強;楊仕明;汪榮泉;房殿春;;食管腺癌組織中核干細(xì)胞因子表達的研究[J];胃腸病學(xué)和肝病學(xué)雜志;2009年06期

9 程鵬;李建生;;不同反流物對十二指腸胃混合食管反流大鼠食管腺癌發(fā)生的影響[J];鄭州大學(xué)學(xué)報(醫(yī)學(xué)版);2011年03期

10 盧嘉臻;王雯;張志堅;李達周;王蓉;;食管腺癌與賁門腺癌的臨床病理特點分析[J];福州總醫(yī)院學(xué)報;2011年02期

相關(guān)會議論文 前1條

1 張遜;David I Watson;Justin R Bessell;;食管腺癌侵及深度及淋巴結(jié)轉(zhuǎn)移與腫瘤預(yù)后的關(guān)系[A];中華醫(yī)學(xué)會第六次全國胸心血管外科學(xué)術(shù)會議論文集（胸外科分冊）[C];2006年

相關(guān)重要報紙文章 前1條

1 武漢同濟醫(yī)院腫瘤中心主任 于世英;胖人更易得癌癥[N];健康報;2008年

相關(guān)博士學(xué)位論文 前1條

1 張濤;選擇性環(huán)氧合酶-2抑制劑塞來昔布用于食管腺癌化學(xué)預(yù)防的實驗研究[D];第四軍醫(yī)大學(xué);2004年

相關(guān)碩士學(xué)位論文 前10條

1 李娜;食管腺癌臨床病理特征及治療模式探討[D];河北醫(yī)科大學(xué);2015年

2 邵順子;重慶地區(qū)食管腺癌和賁門腺癌胃鏡檢出率變化-35年回顧性分析[D];第三軍醫(yī)大學(xué);2015年

3 王恒;食管腺癌組織中CAIX蛋白的表達及其臨床意義[D];鄭州大學(xué);2016年

4 吳敏杰;原發(fā)食管腺癌臨床病理特征及預(yù)后影響因素[D];新鄉(xiāng)醫(yī)學(xué)院;2016年

5 林思捷;Ouabain通過MKK6調(diào)控SOX9基因表達抑制食管腺癌細(xì)胞增殖作用的研究[D];福建醫(yī)科大學(xué);2016年

6 何旭;食管腺癌臨床預(yù)后相關(guān)因素分析[D];河北醫(yī)科大學(xué);2013年

7 穆華;酪酪肽和維生素E琥珀酸酯對人食管腺癌SEG-1細(xì)胞增殖與凋亡的影響[D];河北醫(yī)科大學(xué);2008年

8 魯玉娟;胃泌素及其受體在人食管腺癌細(xì)胞增殖中的作用[D];河北醫(yī)科大學(xué);2008年

9 鄧友君;抑癌基因NDRG2在食管腺癌中的表達及臨床意義[D];遵義醫(yī)學(xué)院;2014年

10 張濤;反流誘發(fā)Barrett’s食管、食管腺癌及其發(fā)展過程中誘導(dǎo)型一氧化氮合酶表達及意義的實驗研究[D];第四軍醫(yī)大學(xué);2001年

，

本文編號：2018706