【作者】 叶广坡；

【导师】 刘弋； 王光升； 曹先东；

【作者基本信息】 安徽医科大学 ， 外科学， 2007， 硕士

【摘要】 背景:大肠癌(colorectal carcinoma,CRC)是消化道常见的恶性肿瘤,国内其发病率在逐年上升。手术切除肿瘤是治疗大肠癌的主要治疗方法,也是唯一可以根治本病的方法,但手术后5年生存率仅有50-60%,原因是对大肠癌的发生、发展仍缺乏深入的认识,早期发现、早期诊断率低,根治性手术后出现复发和转移。除手术治疗外,目前还有化疗、放疗等方法,但疗效有限,副作用大。肿瘤免疫治疗理论上具有靶向杀灭肿瘤细胞,副作用小的特点,是近年来肿瘤治疗的研究热点。因此,寻找一种新的大肠癌肿瘤标志物,研究其在大肠癌中的表达特点和临床意义,有利于深入了解大肠癌的演变过程,提高对本病的认识、诊断和治疗水平,具有重要的意义。上皮细胞粘附分子(epithelial cell adhesion molecule, Ep-CAM)是一种上皮细胞单亚基跨膜糖蛋白;在正常上皮组织呈阴性或弱表达,在癌组织中常过度表达或重新表达;介导Ca2+非依赖性噬同种黏附;可能与癌细胞的增殖、侵袭、去分化及转移相关;是多种上皮恶性肿瘤主动和被动免疫治疗研究的靶分子。目前,国内外未见大肠癌组织与癌旁组织中EP-CAM定量关系以及大肠癌组织EP-CAM表达水平与患者年龄、性别及肿瘤生物学行为关系的文献,此外,其作为一项新的大肠癌肿瘤标志物及免疫治疗靶分子的可行性国内外鲜见报道。目的:(1)研究Ep-CAM在大肠癌组织中的表达水平;(2)了解Ep-CAM表达与大肠癌发生、发展的关系;(3)了解大肠癌组织Ep-CAM表达水平与患者年龄、性别及肿瘤生物学行为的关系;(4)探讨Ep-CAM能否作为大肠癌的肿瘤标志物;(5)比较Ep-CAM与C12作为大肠癌诊断方法的价值;(6)探讨Ep-CAM能否作为大肠癌免疫治疗的靶分子。方法与步骤:(1)分别取53例原发性大肠癌组织和癌旁黏膜组织,制备组织芯片3张并进行免疫组化染色。(2)用形态学图象分析系统测定大肠癌组织和癌旁黏膜组织上皮细胞粘附分子染色积分光密度(IOD of Ep-CAM positive area,以下简称为Ep-CAMIOD)。(3)术前采集大肠癌患者非抗凝血清1ml,用血清蛋白芯片多种肿瘤标志物检测系统(C12)测定12种肿瘤标志物的血清含量。(4)分析大肠癌组织与癌旁组织Ep-CAMIOD之间的关系;大肠癌组织Ep-CAMIOD与患者年龄、性别及肿瘤生物学行为之间的关系。(5)比较C12阳性和阴性组大肠癌组织Ep-CAMIOD之间的差异。结果:(1)大肠癌组织Ep-CAMIOD显著高于癌旁组织Ep-CAMIOD(P<0.01)。(2)大肠癌组织Ep-CAMIOD与患者年龄、性别及肿瘤生物学行为无明显相关性(P>0.05)。(3)50.9﹪的大肠癌患者C12阳性。(4)C12阳性与阴性组大肠癌组织Ep-CAMIOD比较无显著差异(P>0.05)。结论:(1)Ep-CAM参与了大肠癌发生、发展的全过程。(2)大肠癌组织中Ep-CAM表达水平与患者年龄、性别及肿瘤生物学行为无明显相关,是大肠癌发生、发展的一种独立性影响因素。(3)Ep-CAM可能作为大肠癌的肿瘤标志物,可以检出C12阴性病例,如与C12联合检查,将提高对大肠癌的检出率。(4)Ep-CAM可能作为大肠癌主动或被动免疫治疗的靶分子。更多还原

【Abstract】 Background: Colorectal carcinoma (CRC) is a kind of malignant tumor that frequently occurred in gastrointestinal system and its morbidity has been rising in recent years in china. The main treatment to this disease is operation which is the only method for radical cure and only about 50-60% suffers can survival 5 years. At present, it is unknown about the genesis and development of CRC and a few patients was found in the disease early phrase, so that the relapse and metastasis would frequently occurred after operation. Apart from surgical treatment, there are also chemotherapy, radiotherapy and so on for CRC. However, these treatments, generally speaking, have gloomy curative effect and considerable adverse reactions. Immunotherapy for malignant tumors, which, in theory, targets to malignant cells and has low secondary effects, has been exploring for many years. It is very important to find a CRC tumor marker and explore its expresion and clinical value, so that we can make profound apprehension about CRC and elevate the diagnosis ability and therapeutic efficacy for CRC.Epithelial cell adhesion molecule(Ep-CAM)is a transmembrane glycoprotein including one peptide, which was negatively or weakly expressed on norm epithelial cells and frequently overexpressed or re-expressed in epithelial carcinomas. It was reported that the molecule mediates Ca2+-independent homophilic adhesions and may correlate with cellular proliferation, invasion, de-differentiation as well as migration. Now, many kinds of active passive and immunotherapies for several kinds of malignant tumor, which targeted to Ep-CAM ,have been explored.At present, It’s still unknown about about the Ep-CAM quantitive relations between CRC tissue and adjacent mucosa as well as the relations between Ep-CAM level in CRC tissue and patients age, gender as well as tumor biological behaviors of CRC and there are a few studies the possibility to use Ep-CAM as a CRC tumor maker and a immunotherapy target all over the world.Objective: (1)Study EP-CAM expression in malignant tissue of large intestina. (2) Investigate the relations between Ep-CAM expression and CRC genesis and development. (3)Explore the relations between EP-CAM content in colorectal tumor tissue and patients age, gender as well as tumor biological behaviors. (4)Evaluate whether EP-CAM could be used as a tumor marker for CRC. (5) Compare the diognosis value of EP-CAM and C12 for CRC. (6)Evaluate whether EP-CAM could be used as a immunotherapy target for CRC.Methods: (1) 53 pieces of colorectal tumor tissues and the same amount of adjacent mucosa were gathered, then, 3 pieces of tissue micro-array sections were made and immunostained. (2) The integral optical density of Ep-CAM staining area (short for Ep-CAMIOD) was inspected by image analysis morphological system. (3)Before operation, 1ml non-anticoagulation serum was collected from every subject, then12 kinds of tumor markers in serum were detected by 12 tumor markers protein micro-array system. (4)The relations of Ep-CAMIOD between in colorectal tumor tissue and in adjacent mucosa groups were investigated. Malignant tissue group was divided into different sub-groups by patients age, gender and tumor biological behaviors, then, we analyze the relationships of Ep-CAMIOD among sub-groups. (5)We also researched the differences of Ep-CAMIOD between C12 positive and negative groups.Results: (1)The Ep-CAMIOD was significantly higher in CRC group than that in control group(P<0.01). (2)Ep-CAMIOD was not related to patients age, gender and tumor biological behaviors(P>0.05. (3)we found that C12 was positive in 50.9﹪ CRC patients. (4)There was no significant difference of Ep-CAMIOD between C12 positive and negtive groups(P>0.05).Conclusions: (1)Ep-CAM was involved in CRC genesis and development. (2)Ep-CAM expression in cancer tissues had no significant relationships with patients age, gender and tumor biological behaviors and Ep-CAM was a independent inflence factor for CRC genesis and development. (3)Ep-CAM could be used as a tumor marker for CRC, which could found C12 negtive patients, so that, if it was detected together with C12, CRC detection rate may be improved. (4)Ep-CAM may be used as a active or passive immunotherapy target CRC.更多还原