本文選題：E-cadherin + β-catenin��； 參考：《安徽醫(yī)科大學(xué)》2014年碩士論文

【摘要】：研究背景： 宮頸癌是常見的婦科腫瘤，近年來我國每年新增宮頸癌患者13萬余人，約占世界新增病例總數(shù)的28.8%，并逐漸趨于年輕化。若早期被活檢證實，大多數(shù)腫瘤可以完整切除，可以大大降低宮頸癌患者的死亡率及復(fù)發(fā)率。宮頸癌是一個緩慢進展的連續(xù)過程，與患者宮頸人乳頭瘤病毒（human papillomavirus，HPV）感染、人流次數(shù)、性伴侶、初次性交年齡、宮頸腫瘤家族史、產(chǎn)次、地理分布等多種高危因素具有明顯相關(guān)性，約99.7%的宮頸癌均可見HPV感染，HPV感染已成為宮頸癌的最主要病因。在這一過程中，常常伴隨有上皮間質(zhì)轉(zhuǎn)化(epithelial-mesenchymal transition，EMT)現(xiàn)象發(fā)生。 EMT多指上皮細胞在某些特定因子的作用下，細胞連接解體、細胞極性逐漸喪失、細胞骨架重構(gòu)，從而使上皮細胞的極性喪失,遷移和運動能力增強,同時上皮表型丟失而逐漸獲得間質(zhì)表型。E-cadherin是粘附分子中鈣離子依賴的細胞粘附素家族中的一員，在正常上皮的表型的維持中起了關(guān)鍵作用，它的減少或丟失是EMT最重要的標(biāo)志性變化。AKT是PI3K下游最重要的信號分子，它是一種絲氨酸/蘇氨酸激酶，活化的p-AKT通過AKT/GSK-3β/Snail通路誘導(dǎo)EMT的發(fā)生，上皮向間質(zhì)轉(zhuǎn)化，促進惡性腫瘤的侵襲和轉(zhuǎn)移。 子宮頸癌的發(fā)生是一個漸進的過程，由子宮頸上皮內(nèi)瘤變（cervicalintraepithelial neoplasm，CIN），經(jīng)過CIN1、2、3的漸進性，，逐步發(fā)展形成宮頸癌。期間，EMT的發(fā)生與宮頸病變程度及其可能的信號傳導(dǎo)分子還不是很清楚。因此，我們收集不同病變級別的宮頸組織標(biāo)本，分析宮頸組織中E-cadherin表達及AKT磷酸化水平的動態(tài)變化與宮頸病變程度之間的關(guān)系，結(jié)合相關(guān)信號分子及宮頸上皮分化指標(biāo)的檢測，進一步探討其可能的分子學(xué)機制，為完善宮頸癌發(fā)病機制及臨床診治提供實驗室證據(jù)。 目的：分析E-cadherin表達和AKT磷酸化水平與宮頸病變程度的相關(guān)性，探討EMT與疾病進展的關(guān)系及其可能的信號通路。 方法：收集20例正常宮頸組織、30例CIN組織、50例宮頸鱗癌組織，采用實時熒光定量PCR、免疫組化和WB分別檢測組織中E-cadherin、β-catenin和Loricrin的mRNA及蛋白表達情況，同時檢測不同級別病變組織中AKT磷酸化程度的變化；分析E-cadherin表達和AKT磷酸化水平的相關(guān)性，及它們與子宮頸病變程度和宮頸鱗癌分化程度的相關(guān)性。 結(jié)果：（1）免疫組化結(jié)果顯示，隨著宮頸病變的進展，E-cadherin、Loricrin的表達逐漸降低，甚至缺失（P＜0.05）；在正常宮頸上皮β-catenin表達于胞膜，隨著宮頸病變的進展，逐漸由胞膜向胞漿和胞核轉(zhuǎn)移，且表達陽性率逐漸降低（P＜0.05）；p-AKT的表達則隨病變進展而逐漸升高（P＜0.05）。 （2）WB結(jié)果顯示，隨著宮頸病變的進展，E-cadherin、Loricrin、β-catenin的表達逐漸降低，甚至缺失（P＜0.05）；p-AKT的表達則隨病變進展而逐漸升高（P＜0.05），與免疫組化結(jié)果一致。 （3）定量PCR結(jié)果亦證實，隨著宮頸病變的進展，E-cadherin、Loricrin、β-catenin的mRNA表達水平逐漸降低，與蛋白表達結(jié)果一致（P值均＜0.05）。 （4）在宮頸癌中，免疫組化和WB結(jié)果顯示，隨著腫瘤組織分化程度的降低，E-cadherin、Loricrin、β-catenin的表達逐漸降低，甚至缺失（P＜0.05），而AKT的磷酸化水平則呈現(xiàn)逐漸升高的趨勢（P＜0.05）。 （5）相關(guān)分析顯示，E-cadherin的表達與宮頸病變的程度呈負相關(guān)（rs=-.688， P＜0.01），而p-AKT的表達則呈正相關(guān)（rs=.462，P＜0.01）。E-cadherin的表達與鱗癌的分化程度呈正相關(guān)(rs=.438，P＜0.01）而p-AKT的表達則呈負相關(guān)（rs=-.692, P＜0.01）。 （7）E-cadherin表達與AKT磷酸化程度呈負相關(guān)(rs=.828, P＜0.01)。 結(jié)論： （1）E-cadherin表達缺失和AKT磷酸化水平增高與宮頸病變的進展密切相關(guān)； （2）β-catenin細胞內(nèi)異常定位及AKT異�；罨趯m頸上皮EMT發(fā)揮關(guān)鍵作用； （3）E-cadherin和p-AKT可用于監(jiān)測宮頸病變的進展，指導(dǎo)臨床治療及預(yù)后判斷。
[Abstract]:Research background:
Cervical cancer is a common gynecologic tumor. In recent years, more than 13 thousands of new cervical cancer patients have been added, accounting for about 28.8% of the total number of new cases in the world, and gradually tend to be younger. If early biopsy proved that most of the tumors can be completely removed, the mortality and recurrence rate of cervical cancer patients can be greatly reduced. Cervical cancer is a slow progress. The continuous process, with the patient's cervical human papillomavirus (human papillomavirus, HPV) infection, the number of people, sexual partners, the age of first sexual intercourse, the family history of cervical cancer, birth, geographical distribution and other high-risk factors, about 99.7% of cervical cancer can be found to be HPV infection, HPV infection has become the main cause of cervical cancer. During the process, epithelial-mesenchymal transition (EMT) often occurs.
In EMT, the epithelial cells are disintegrated, the cell polarity is gradually lost and the cytoskeleton reconstructs, so that the epithelia is lost, the migration and movement ability is enhanced, and the epithelial phenotype is lost and the.E-cadherin is a calcium dependent cell adhesion family in the adhesion molecule. A member of the family, which plays a key role in the maintenance of normal epithelial phenotype, its reduction or loss is the most important symbolic change in EMT.AKT, the most important signal molecule in the lower reaches of PI3K. It is a serine / threonine kinase, the activated p-AKT induces the occurrence of EMT through the AKT/GSK-3 beta /Snail pathway, the epithelial transformation to the interstitial, and the promotion of evil. The invasion and metastasis of sexual tumors.
The occurrence of cervical cancer is a gradual process, from the cervicalintraepithelial neoplasm (CIN) to the gradual development of CIN1,2,3 to form cervical cancer. During the period, the occurrence of EMT and the degree of cervical lesions and the possible signal transduction molecules are not clear. Therefore, we collect different pathological grades. To analyze the relationship between the dynamic changes of E-cadherin and the level of AKT phosphorylation in cervical tissue and the degree of cervical lesions, and to further explore the possible molecular mechanism of the cervical tissue, and provide laboratory evidence for the improvement of the pathogenesis and clinical diagnosis and treatment of cervical cancer.
Objective: to analyze the correlation between E-cadherin expression and AKT phosphorylation level and the degree of cervical lesions, and to explore the relationship between EMT and disease progression and its possible signaling pathways.
Methods: 20 normal cervical tissues, 30 CIN tissues and 50 cases of cervical squamous cell carcinoma were detected by real-time fluorescence quantitative PCR. Immunohistochemistry and WB were used to detect the mRNA and protein expression of E-cadherin, beta -catenin and Loricrin in tissues. The changes in the degree of AKT phosphorylation in different pathological tissues were detected, and the expression of E-cadherin was analyzed. The correlation of AKT phosphorylation level and their correlation with the degree of cervical lesion and the degree of differentiation of cervical squamous cell carcinoma.
Results: (1) the immunohistochemical results showed that with the progression of cervical lesions, the expression of E-cadherin and Loricrin decreased gradually and even lost (P < 0.05); in normal cervical epithelium, the expression of beta -catenin in the cell membrane was gradually transferred from the membrane to the cytoplasm and nucleus, and the positive rate of the expression gradually decreased (P < 0.05); p-AKT The expression was gradually increased with the progression of the lesion (P < 0.05).
(2) WB results showed that with the progression of cervical lesions, the expression of E-cadherin, Loricrin, and beta -catenin gradually decreased and even lost (P < 0.05), and the expression of p-AKT increased gradually with the progression of the lesion (P < 0.05), which was consistent with the immunohistochemical results.
(3) quantitative PCR results also confirmed that with the progression of cervical lesions, the mRNA expression level of E-cadherin, Loricrin, and beta -catenin decreased gradually, and was consistent with the protein expression results (P value < 0.05).
(4) in cervical cancer, immunohistochemical and WB results showed that the expression of E-cadherin, Loricrin, and beta -catenin decreased gradually and even disappeared (P < 0.05), and the level of phosphorylation of AKT was gradually increasing with the decrease of the degree of differentiation of tumor tissue (P < 0.05).
(5) the correlation analysis showed that the expression of E-cadherin was negatively correlated with the degree of cervical lesions (rs=-.688, P < 0.01), while the expression of p-AKT was positively correlated (rs=.462, P < 0.01) and the expression of.E-cadherin was positively correlated with the degree of squamous cell differentiation (rs=.438, P < 0.01) and the expression of p-AKT was negative correlation (rs=-.692, P < 0.01).
(7) the expression of E-cadherin was negatively correlated with the degree of AKT phosphorylation (rs=.828, P < 0.01).
Conclusion:
(1) increased expression of E-cadherin and increased phosphorylation of AKT are closely related to the progression of cervical lesions.
(2) abnormal localization of -catenin cells and abnormal activation of AKT play a key role in cervical EMT.
(3) E-cadherin and p-AKT can be used to monitor the progress of cervical lesions and guide clinical treatment and prognosis.

【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R737.33

【參考文獻】

相關(guān)期刊論文 前1條

1 ;Caveolin-1,E-cadherin and β-catenin in Gastric Carcinoma,Precancerous Tissues and Chronic Non-atrophic Gastritis[J];Chinese Journal of Cancer Research;2012年01期

本文編號：1808638