Six1、FOXC2及E-cad在结肠癌中的表达及临床意义

发布时间：2018-05-29 19:31

本文选题：结肠恶性肿瘤 + Six1　；参考：《承德医学院》2017年硕士论文

【摘要】：纵观消化道环球发病率最高的几种恶性疾病中结肠癌的占有比率愈来愈高。结肠癌在我国的发病率及死亡率亦较前大幅增长。以往众多研究结论显示,结肠癌患者治愈率降低、死亡率升高的关键因素是发生转移,而结肠癌的发生发展是包含多个阶段涉及很多因素的综合复杂病理过程,防治结肠癌的转移成为医务工作者一直以来较难攻克的难题。然而,随着上皮间质细胞转化(EMT)的发现及其在恶性肿瘤中的深入研究,提示包括结肠癌在内许多肿瘤细胞浸润、早期转移过程中发生了EMT,因此,通过探索结肠癌EMT相关分子机制,为提高结肠癌患者生存率及改善预后提供较为可能的有效途径。同源盒基因1(Six1)可与DNA特定部位结合,特异性调节下游基因的表达,为同源盒基因家族成员,在细胞增殖、分化、黏着、转移等重要过程作用关键。EMT是30多年前已经被提出的一个分子事件,它是指上皮细胞在某些病理或生理条件下转变成具有移动能力的间质细胞的过程。大量的研究已经表明,其与大部分恶性肿瘤的原发性浸润和继发性转移密不可分。随着研究的不断深入,越来越多EMT相关的关键调节因子和通路被不断发现和完善。Six1蛋白是近年来研究EMT相关分子的热点转录调节因子之一,目前已有研究提示,细胞周期的调控、胚胎生长和器官成熟、癌细胞的激活及转移的推动等过程都与Six1有关。其在肿瘤细胞中高表达,通过诱导细胞增生、抑制凋亡,甚至为恶性细胞创造EMT发生条件而促进转移等过程,参与肿瘤发生发展。叉头框蛋白C2(Forkhead box protein C2,FOXC2),也被称为间充质叉头蛋白1(Mesenchyme fork head protein 1,MFHl),与叉头框家族其他成员一样具有特定DNA结合域,参与DNA转录过程。研究发现,FOXC2是EMT的重要调节者之一,其他某些EMT相关转录因子受FOXC2诱导表达,同时FOXC2也能正向提高自身在上皮细胞中的表达,而且还可上调基质金属蛋白酶2(MMP-2)和基质金属蛋白酶9(MMP-9)的水平,促进间质分化及细胞的移动,进而为协助肿瘤细胞在临近组织中的浸润和远处脏器的转移提供基础。E-钙黏蛋白(E-cad)遍及各种上皮细胞表面分布,起到促进细胞间相互衔接粘着功能,是保持细胞极性、维持细胞形态的结构分子,同时也是介导上皮细胞之间及其与基质间通信的媒介。E-cad缺失表达或功能缺损后上皮细胞可经历改变极性、降低黏附力等过程,最终呈现非上皮特征。经研究发现,在上皮细胞发生EMT过程中,E-cad表达下调是普遍现象,它的表达缺失在细胞恶性表型的转变中具有因果关系,所以E-cad的表达下调是EMT的最显著性的特征之一。目的:通过检测Six1、FOXC2及E-cad在结肠癌及癌旁正常结肠组织中的表达水平,剖析结肠癌中Six1、FOXC2及E-cad的表达水平与临床病理特征及在肿瘤增殖发展过程中彼此之间的相关性。Six1、FOXC2为肿瘤发生发展治疗新靶点提供新的理论基础以及联合检测三者为判断临床预后提供更多的依据。方法:收集病理明确诊断为结肠腺癌的蜡块固定组织70例和癌旁正常结肠蜡块固定组织35例,术前患者均无实行放疗、化疗等任何新辅助治疗。用免疫组织化学法检测结肠癌组织中Six1、FOXC2及E-cad表达水平,分析性别、年龄、肿瘤大小、浸润深度及淋巴结转移与它们是否存在相关性。数据分析使用SPSS 19.0,χ2检验统计计数资料,Spearman秩相关分析判断指标间相关性,检验标准为α=0.05,以P0.05为差异有统计学意义。结果:免疫组织化学结果1.Six1、FOXC2及E-cad在结肠癌组织和癌旁正常结肠组织中都有不同程度表达。Six1和FOXC2主要于细胞质染色阳性,在癌组织中的阳性表达率分别为82.9%(58/70)和74.3%(52/70),均显著高于癌旁正常组织中的阳性表达率20.0%(7/35)和22.9%(8/35),差异均具有统计学意义(χ2=39.092,P=0.000;χ2=25.200,P=0.000)。E-cad主要于细胞膜染色阳性,在结肠癌组织表达阳性率为21.4%(15/70),与正常结肠组织中的表达阳性率80.0%(28/35)比显著降低,差异有统计学意义(χ2=33.103,P=0.000)。2.Six1、FOXC2及E-cad于结肠癌中的表达水平与性别、年龄、肿瘤大小、分化程度无关联,且相关性无统计学差异(P0.05),而与TNM分期、浸润深度及淋巴结转移具有相关性,且具有统计学意义(P0.05)。3.在结肠癌中Six1与FOXC2的阳性表达呈正相关(r=0.426,P0.01);E-cad的阳性表达分别与Six1、FOXC2呈负相关(r=-0.480,P0.05;r=-0.728,P0.05),差异均具有统计学意义(P0.05)。结论:Six1、FOXC2在结肠癌组织中的阳性率均明显高于癌旁正常结肠组织,而E-cad在癌组织的阳性率显著降低,且三者在恶性组织中的表达均与肿瘤浸润深度、淋巴结转移、TNM分期有关。侵袭管壁越深,有淋巴结转移,TNM分期越晚,则Six1、FOXC2阳性率越高,E-cad阳性率越低,提示三者与结肠癌的恶性行为及预后判断可能存在相关性,可较为敏感的提示癌细胞发生浸润转移。Six1和FOXC2的阳性表达呈正相关,且二者均与E-cad呈负相关。而EMT会导致肿瘤浸润转移,故联合检测三者的表达水平可能会在结肠癌EMT治疗新靶点方面提供理论依据,及进一步探讨其判断结肠癌患者的临床预后提供一定的研究基础。
[Abstract]:The incidence of colon cancer in several malignant diseases with the highest incidence of global digestive tract is higher and higher. The incidence and mortality of colon cancer in China have also increased significantly. The complex pathological process involving many factors is involved in many factors, and the prevention and treatment of colon cancer has been a difficult problem for medical workers. However, with the discovery of epithelial mesenchymal cell transformation (EMT) and its deep study in malignant tumor, it suggests that many tumor cells including colon cancer, including the infiltration of cancer cells, and early metastasis. EMT has occurred in the process. Therefore, by exploring the EMT related molecular mechanism of colon cancer, it provides a more effective way to improve the survival rate and improve the prognosis of colon cancer patients. Homologous box gene 1 (Six1) can be combined with specific sites of DNA, specifically regulate the expression of downstream genes, for the family members of the homologous gene family, cell proliferation, differentiation, and adhesion. The key.EMT is a molecular event that has been proposed more than 30 years ago. It refers to the process of epithelial cells in some pathological or physiological conditions into a cell with a mobile capacity. A large number of studies have shown that the primary invasion and secondary metastasis of most malignant tumors are incompatible. In recent years, more and more key regulatory factors and pathways related to EMT have been discovered and perfected by more and more.Six1 proteins, which are one of the hot transcriptional regulators for the study of EMT related molecules in recent years. There have been some research on the regulation of cell cycle, embryo growth and organ maturation, the promotion of cancer cell activation and metastasis. The process is related to Six1, which is highly expressed in tumor cells, and is involved in the development of tumor by inducing cell proliferation, inhibiting apoptosis and even creating EMT conditions for malignant cells, and involved in the development of tumor. Forkhead box protein C2 (FOXC2), also known as mesenchymal fork head protein 1 (Mesenchyme fork head protein 1,) MFHl), as with other members of the forked frame family, has a specific DNA binding domain and participates in the DNA transcriptional process. It is found that FOXC2 is one of the important regulators of EMT. Some of the other EMT related transcription factors are induced by FOXC2, and FOXC2 can positively increase its expression in the epithelial cells, and also up regulation of matrix metalloproteinase 2 (MMP). -2) and matrix metalloproteinase 9 (MMP-9) levels, promoting interstitial differentiation and cell migration, and providing the basis for assisting tumor cells to infiltrate in adjacent tissues and metastasis of distant organs to provide the basis for the distribution of.E- calcium mucin (E-cad) throughout the surface of epithelial cells, promoting intercellular adhesion and adhesion, and maintaining cell polarity. The structural molecules that maintain cell morphology are also a medium for mediating and communicating between the epithelial cells and the matrix between the medium.E-cad and the loss of function after the deletion of the epithelial cells. The epithelial cells can undergo the process of changing polarity, reducing adhesion and other processes, and eventually showing non epithelial characteristics. The study found that the down regulation of E-cad expression in the EMT process of epithelial cells is common. The expression of E-cad is one of the most significant characteristics of EMT. Objective: to analyze the expression level of Six1, FOXC2 and E-cad in colon and adjacent normal colonic tissues, and to analyze the expression level and clinical level of Six1, FOXC2 and E-cad in colon cancer. The pathological features and the correlation between each other in the process of tumor proliferation and development.Six1, FOXC2 provides a new theoretical basis for the new target of tumor development and development, and the joint detection three provides more basis for judging the clinical prognosis. Methods: 70 cases of paraffin fixed tissue and normal paracancerous junctions of colon adenocarcinoma were diagnosed by collecting pathology. 35 cases of intestinal wax block fixed tissue, no new adjuvant therapy, such as radiotherapy and chemotherapy, were used before the operation. The expression of Six1, FOXC2 and E-cad in colon cancer tissue was detected by immunohistochemistry. The correlation between sex, age, tumor size, depth of invasion and lymph node metastasis was correlated with them. Data analysis used SPSS 19, chi 2 test Statistical count data, Spearman rank correlation analysis was used to determine the correlation between the indexes, the test standard was alpha =0.05, and the difference between P0.05 and 1.Six1, FOXC2 and E-cad in colon cancer tissues and normal colon tissues were different to some extent,.Six1 and FOXC2 were mainly positive for cytoplasmic staining and in cancer. The positive expression rates in the tissues were 82.9% (58/70) and 74.3% (52/70), respectively, which were significantly higher than those in the normal tissues by 20% (7/35) and 22.9% (8/35). The difference was statistically significant (x 2=39.092, P=0.000; Chi 2=25.200, P=0.000).E-cad mainly in cell membrane staining, and the positive rate in colon cancer tissue was 21.4% (15/70). The positive rate of expression of 80% (28/35) in the normal colonic tissue was significantly lower than that in the colon (x 2=33.103, P=0.000).2.Six1. The expression level of FOXC2 and E-cad in colon cancer was not associated with sex, age, tumor size, and degree of differentiation, and the correlation was not statistically significant (P0.05), but with TNM staging, infiltration depth and lymph node transfer. The positive expression of Six1 and FOXC2 in colon cancer was positively correlated (r=0.426, P0.01) in colon cancer (r=0.426, P0.01), and the positive expression of E-cad was negatively correlated with Six1 and FOXC2 respectively (r=-0.480, P0.05; r=-0.728, Six1). Conclusion: the positive rate of E-cad in colon cancer tissues is all The positive rate of E-cad in the normal colon was significantly higher than that in the cancer tissue, and the expression of the three in the malignant tissue was significantly lower, and the expression of the three in the malignant tissue was related to the depth of tumor infiltration, lymph node metastasis and TNM staging. The deeper the invasion tube wall, the metastasis of the lymph nodes, the later the TNM staging, the higher the positive rate of FOXC2, the lower the positive rate of E-cad, suggesting the three and the knot. There may be a correlation between the malignant behavior and prognosis of colorectal cancer. It is more sensitive to suggest that the positive expression of.Six1 and FOXC2 is positively correlated with the invasion and metastasis of cancer cells, and all of the two are negatively correlated with E-cad. And EMT may lead to tumor invasion and metastasis, so the expression level of the combined detection of the three may be in the new target of colon cancer EMT treatment. It provides a theoretical basis for further study of its clinical prognosis in colon cancer patients.
【学位授予单位】：承德医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.35

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