E-cadherin表达和AKT磷酸化水平与宫颈病变程度的相关性分析

发布时间：2018-04-27 01:39

本文选题：E-cadherin + β-catenin　；参考：《安徽医科大学》2014年硕士论文

【摘要】：研究背景：宫颈癌是常见的妇科肿瘤，近年来我国每年新增宫颈癌患者13万余人，约占世界新增病例总数的28.8%，并逐渐趋于年轻化。若早期被活检证实，大多数肿瘤可以完整切除，可以大大降低宫颈癌患者的死亡率及复发率。宫颈癌是一个缓慢进展的连续过程，与患者宫颈人乳头瘤病毒（human papillomavirus，HPV）感染、人流次数、性伴侣、初次性交年龄、宫颈肿瘤家族史、产次、地理分布等多种高危因素具有明显相关性，约99.7%的宫颈癌均可见HPV感染，HPV感染已成为宫颈癌的最主要病因。在这一过程中，常常伴随有上皮间质转化(epithelial-mesenchymal transition，EMT)现象发生。 EMT多指上皮细胞在某些特定因子的作用下，细胞连接解体、细胞极性逐渐丧失、细胞骨架重构，从而使上皮细胞的极性丧失,迁移和运动能力增强,同时上皮表型丢失而逐渐获得间质表型。E-cadherin是粘附分子中钙离子依赖的细胞粘附素家族中的一员，在正常上皮的表型的维持中起了关键作用，它的减少或丢失是EMT最重要的标志性变化。AKT是PI3K下游最重要的信号分子，它是一种丝氨酸/苏氨酸激酶，活化的p-AKT通过AKT/GSK-3β/Snail通路诱导EMT的发生，上皮向间质转化，促进恶性肿瘤的侵袭和转移。子宫颈癌的发生是一个渐进的过程，由子宫颈上皮内瘤变（cervicalintraepithelial neoplasm，CIN），经过CIN1、2、3的渐进性，，逐步发展形成宫颈癌。期间，EMT的发生与宫颈病变程度及其可能的信号传导分子还不是很清楚。因此，我们收集不同病变级别的宫颈组织标本，分析宫颈组织中E-cadherin表达及AKT磷酸化水平的动态变化与宫颈病变程度之间的关系，结合相关信号分子及宫颈上皮分化指标的检测，进一步探讨其可能的分子学机制，为完善宫颈癌发病机制及临床诊治提供实验室证据。目的：分析E-cadherin表达和AKT磷酸化水平与宫颈病变程度的相关性，探讨EMT与疾病进展的关系及其可能的信号通路。方法：收集20例正常宫颈组织、30例CIN组织、50例宫颈鳞癌组织，采用实时荧光定量PCR、免疫组化和WB分别检测组织中E-cadherin、β-catenin和Loricrin的mRNA及蛋白表达情况，同时检测不同级别病变组织中AKT磷酸化程度的变化；分析E-cadherin表达和AKT磷酸化水平的相关性，及它们与子宫颈病变程度和宫颈鳞癌分化程度的相关性。结果：（1）免疫组化结果显示，随着宫颈病变的进展，E-cadherin、Loricrin的表达逐渐降低，甚至缺失（P＜0.05）；在正常宫颈上皮β-catenin表达于胞膜，随着宫颈病变的进展，逐渐由胞膜向胞浆和胞核转移，且表达阳性率逐渐降低（P＜0.05）；p-AKT的表达则随病变进展而逐渐升高（P＜0.05）。（2）WB结果显示，随着宫颈病变的进展，E-cadherin、Loricrin、β-catenin的表达逐渐降低，甚至缺失（P＜0.05）；p-AKT的表达则随病变进展而逐渐升高（P＜0.05），与免疫组化结果一致。（3）定量PCR结果亦证实，随着宫颈病变的进展，E-cadherin、Loricrin、β-catenin的mRNA表达水平逐渐降低，与蛋白表达结果一致（P值均＜0.05）。（4）在宫颈癌中，免疫组化和WB结果显示，随着肿瘤组织分化程度的降低，E-cadherin、Loricrin、β-catenin的表达逐渐降低，甚至缺失（P＜0.05），而AKT的磷酸化水平则呈现逐渐升高的趋势（P＜0.05）。（5）相关分析显示，E-cadherin的表达与宫颈病变的程度呈负相关（rs=-.688， P＜0.01），而p-AKT的表达则呈正相关（rs=.462，P＜0.01）。E-cadherin的表达与鳞癌的分化程度呈正相关(rs=.438，P＜0.01）而p-AKT的表达则呈负相关（rs=-.692, P＜0.01）。（7）E-cadherin表达与AKT磷酸化程度呈负相关(rs=.828, P＜0.01)。结论：（1）E-cadherin表达缺失和AKT磷酸化水平增高与宫颈病变的进展密切相关；（2）β-catenin细胞内异常定位及AKT异常活化在宫颈上皮EMT发挥关键作用；（3）E-cadherin和p-AKT可用于监测宫颈病变的进展，指导临床治疗及预后判断。
[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.

【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.33

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