發(fā)布時(shí)間：2018-06-07 08:38

  本文選題：食管鱗癌 + Smac　； 參考：《山東大學(xué)》2017年碩士論文

【摘要】：[目的與背景]食管癌是最常見(jiàn)的惡性腫瘤之一,其發(fā)病率在全世界惡性腫瘤中居第9位,在我國(guó)居第5位,其死亡率在全球范圍內(nèi)居第8位,在我國(guó)居第4位。我國(guó)食管癌以食管鱗狀細(xì)胞癌為主(簡(jiǎn)稱食管鱗癌,esophageal squamous cell carcinoma,ESCC),占食管癌的90%以上。早期食管癌手術(shù)切除后5年生存率可達(dá)85%-90%,但因其缺乏典型的臨床癥狀而難以早期診斷治療。大部分患者在出現(xiàn)吞咽困難或轉(zhuǎn)移癥狀時(shí)才去就診,此時(shí)多已經(jīng)處于中晚期,5年生存率僅為60%-15%。為了改善食管癌的預(yù)后,在評(píng)估及診療方面的分子技術(shù)尚有待改進(jìn)。由于癌細(xì)胞通常具有逃避凋亡的進(jìn)化機(jī)制,因此與細(xì)胞凋亡途徑相接合的診療手段充滿前景。凋亡抑制因子(inhibitor of apoptosis proteins,IAPs)阻礙了細(xì)胞凋亡,這與腫瘤侵襲、進(jìn)展、治療失敗和預(yù)后不良有關(guān)。腫瘤細(xì)胞高表達(dá)IAPs是導(dǎo)致腫瘤細(xì)胞抵抗凋亡的關(guān)鍵。人類(lèi)第二線粒體源性胱氨酸酶激活因子(Second mitochondria-derived activator of caspase,Smac)是凋亡途徑中的一種重要蛋白,通過(guò)消除IPAs對(duì)胱天蛋白酶(cysteine aspartate-specific proteases,caspase)的抑制發(fā)揮促凋亡作用,因此Smac被認(rèn)為是食管癌治療的潛在靶點(diǎn)之一。闡明Smac在食管鱗狀細(xì)胞癌侵襲中的表達(dá)情況、生物學(xué)功能及臨床意義可能具有重要的價(jià)值。本課題通過(guò)免疫組織化學(xué)方法檢測(cè)Smac在食管鱗癌組織、癌旁組織及正常食管組織中的表達(dá)情況,探討其與臨床病理特點(diǎn)之間的關(guān)系,并通過(guò)轉(zhuǎn)染上調(diào)Smac在食管鱗癌細(xì)胞ECA109中的表達(dá),驗(yàn)證Smac對(duì)食管鱗癌細(xì)胞體外侵襲行為的調(diào)控作用。[材料與方法]收集山大二院2015年1月至2016年8月期間手術(shù)切除的食管鱗癌組織、癌旁組織和正常食管組織共68組,常規(guī)石蠟包埋切片,通過(guò)免疫組織化學(xué)方法檢測(cè)Smac食管鱗癌組織及對(duì)應(yīng)癌旁組織和正常組織中的表達(dá)情況,運(yùn)用X2檢驗(yàn)分析Smac的表達(dá)與食管鱗癌患者臨床病理參數(shù)的關(guān)系。通過(guò)qRT-PCR獲取Smac蛋白的cDNA,從瓊脂糖凝膠中回收靶片段,構(gòu)建含Smac基因片段的真核表達(dá)載體pcDNA3.1-Smac,通過(guò)瞬時(shí)轉(zhuǎn)染技術(shù)獲得穩(wěn)定高表達(dá)Smac的食管鱗癌細(xì)胞株 ECA109/pcDNA3.1-Smac(ECA109/Smac 組)和對(duì)照組 ECA109/pcDNA3.1(ECA109/對(duì)照組),并設(shè)立空白對(duì)照(ECA109組)。采用Western blotting技術(shù)驗(yàn)證Smac蛋白水平的過(guò)表達(dá)率。采用transwell小室檢測(cè)Smac對(duì)食管鱗癌細(xì)胞體外侵襲能力的調(diào)控作用。[實(shí)驗(yàn)結(jié)果]根據(jù)免疫組化結(jié)果,食管鱗癌組織中的Smac陽(yáng)性率為54.41%,明顯低于癌旁組織和正常食管組織Smac的表達(dá)率(分別為89.71%,92.65%;P0.05),Smac的表達(dá)水平與食管鱗癌的TNM分期、浸潤(rùn)深度、淋巴結(jié)轉(zhuǎn)移情況顯著相關(guān)(P0.05),與年齡、性別、分化程度無(wú)關(guān)(P0.05)。Western blotting顯示,ECA109/Smac組的Smac蛋白表達(dá)升高。Transwell侵襲實(shí)驗(yàn)表明與ECA109/對(duì)照組和ECA109組相比,ECA109/Smac組的細(xì)胞侵襲能力降低(P＜0.05)。[實(shí)驗(yàn)結(jié)論]食管鱗癌組織中的Smac明顯降低,上調(diào)Smac的表達(dá)后,食管鱗癌細(xì)胞的侵襲能力降低,表明Smmac在食管鱗癌的發(fā)生發(fā)展及轉(zhuǎn)移中發(fā)揮重要的作用,進(jìn)一步研究Smac的生物學(xué)特性可能有益于治療食管癌。
[Abstract]:[Objective and background] cancer of the esophagus is one of the most common malignant tumors. Its incidence is the ninth most malignant tumor in the world and the fifth place in our country. The mortality rate is the eighth in the world and the fourth in China. Esophageal squamous cell carcinoma is mainly esophageal squamous cell carcinoma (esophageal squamous cell carcinoma, ESCC), More than 90% of the cancer of the esophagus is accounted for. The 5 year survival rate of early esophageal cancer is up to 85%-90%, but it is difficult to be diagnosed early because of its lack of typical clinical symptoms. Most patients go to the doctor when they have dysphagia or metastases. At this time, most of them are in the middle and late stages, and the 5 year survival rate is only 60%-15%. in order to improve the precondition of esophageal cancer. Molecular techniques in evaluation and diagnosis have yet to be improved. Because cancer cells usually have an evolutionary mechanism to escape apoptosis, the diagnostic and therapeutic methods that join the apoptotic pathway are full of prospects. Apoptosis inhibitor (inhibitor of apoptosis proteins, IAPs) hinders apoptosis, which is associated with tumor invasion, progress, treatment failure and The high expression of IAPs in tumor cells is the key to the resistance to apoptosis of tumor cells. Human second mitochondrial Second mitochondria-derived activator of caspase (Smac) is an important protein in the apoptotic pathway, by eliminating IPAs to cystine (cysteine aspartate-specific Pro). The inhibition of teases, caspase) plays a role in promoting apoptosis, so Smac is considered to be one of the potential targets for the treatment of esophageal cancer. It is possible to clarify the expression, biological function and clinical significance of Smac in the invasion of squamous cell carcinoma of the esophagus. This topic is to detect Smac in the tissue of esophageal squamous cell carcinoma and the para cancer by immunohistochemical method. The relationship between the expression of tissue and normal esophageal tissue and its relationship with the clinicopathological characteristics, and the expression of Smac in ECA109 of esophageal squamous cell carcinoma cells by transfection, and to verify the regulation of Smac on the invasion of esophageal squamous cell carcinoma cells in vitro. [materials and methods] collect the operation cut from January 2015 to August 2016. 68 groups of esophageal squamous cell carcinoma, para cancerous tissue and normal esophageal tissue were divided into 68 groups. Routine paraffin embedded sections were used to detect the expression of Smac esophageal squamous cell carcinoma tissue and its para cancerous tissue and normal tissues by immunohistochemistry. The relationship between the expression of Smac and the clinicopathological parameters of the patients with esophageal squamous cell carcinoma was analyzed by X2 test. R obtained the cDNA of Smac protein, recovered the target fragment from agarose gel and constructed the eukaryotic expression vector containing the Smac gene fragment, pcDNA3.1-Smac, and obtained the ECA109/pcDNA3.1-Smac (ECA109/Smac group) and the control group ECA109/pcDNA3.1 (ECA109/ control group), which stabilized the high expression of Smac by transient transfection, and set up a blank pair. Western blotting technique was used to verify the overexpression of Smac protein level. The regulation of Smac on the invasion ability of esophageal squamous cell carcinoma cells was detected by Transwell chamber. [experimental results] the positive rate of Smac in esophageal squamous cell carcinoma was 54.41% according to the immunohistochemical results, which was significantly lower than that of the Para cancerous tissue and the normal esophageal tissue S. The expression rate of MAC (89.71%, 92.65%; P0.05). The expression level of Smac was significantly correlated with the TNM staging, infiltration depth and lymph node metastasis of esophageal squamous cell carcinoma (P0.05). It was not related to age, sex, differentiation degree (P0.05).Western blotting, the increase of Smac protein expression in the ECA109/Smac group showed that.Transwell invasion experiment showed that the ECA109/ control group was in the ECA109/ control group. Compared with the ECA109 group, the cell invasion ability of ECA109/Smac group decreased (P < 0.05). [experimental conclusions] the Smac in esophageal squamous cell carcinoma decreased obviously. After the expression of Smac, the invasion ability of esophageal squamous cell carcinoma cells decreased, indicating that Smmac plays an important role in the development and metastasis of esophageal squamous cell carcinoma, and further studies the biological characteristics of Smac. Sex may be beneficial to the treatment of cancer of the esophagus.
【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級(jí)別】：碩士
【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8條

1 陳萬(wàn)青;鄭榮壽;張思維;曾紅梅;鄒小農(nóng);赫捷;;2013年中國(guó)惡性腫瘤發(fā)病和死亡分析[J];中國(guó)腫瘤;2017年01期

2 李鵬;陳光勇;王擁軍;許昌芹;;中國(guó)早期食管鱗狀細(xì)胞癌及癌前病變篩查與診治共識(shí)(2015年·北京)[J];中國(guó)醫(yī)刊;2016年01期

3 赫捷;邵康;;中國(guó)食管癌流行病學(xué)現(xiàn)狀、診療現(xiàn)狀及未來(lái)對(duì)策[J];中國(guó)癌癥雜志;2011年07期

4 張小剛;鐘理;王建飛;;食管癌危險(xiǎn)因素及預(yù)防研究進(jìn)展[J];世界華人消化雜志;2009年07期

5 李?lèi)?ài)光;孟憲利;王洪琰;劉慶熠;何明;王世杰;;食管鱗癌組織中Smac、survivin的表達(dá)及其臨床意義[J];山東醫(yī)藥;2008年36期

6 ;Relationship between expression of Smac and Survivin and apoptosis of primary hepatocellular carcinoma[J];Hepatobiliary & Pancreatic Diseases International;2006年04期

7 王國(guó)清,喬友林;癌前病變研究——控制食管癌發(fā)展的關(guān)鍵[J];腫瘤研究與臨床;2003年01期

8 王國(guó)清;食管癌高發(fā)現(xiàn)場(chǎng)早診早治 30年臨床研究經(jīng)驗(yàn)[J];中國(guó)醫(yī)學(xué)科學(xué)院學(xué)報(bào);2001年01期

相關(guān)碩士學(xué)位論文 前1條

1 陳鳳娟;Smac在青海地區(qū)食管鱗癌中的表達(dá)及臨床意義[D];青海大學(xué);2014年

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