[1]陈曦,黄悦,于彬,等.结直肠癌单克隆抗体的制备及生物学特性鉴定[J].陆军军医大学学报(原第三军医大学学报),2007,29(10):896-898.
 CHEN Xi,HUANG Yue,YU Bin,et al.Preparing and identifying a monoclonal antibody against human colorectal carcinoma[J].J Amry Med Univ (J Third Mil Med Univ),2007,29(10):896-898.
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结直肠癌单克隆抗体的制备及生物学特性鉴定
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
29卷
期数:
2007年第10期
页码:
896-898
栏目:
论著
出版日期:
2007-05-30

文章信息/Info

Title:
Preparing and identifying a monoclonal antibody against human colorectal carcinoma
作者:
陈曦黄悦于彬梅峰韩鹃周德山
第三军医大学基础医学部组织胚胎学教研室,重庆市神经科学研究所
Author(s):
CHEN Xi HUANG Yue YU Bin MEI Feng HAN Juan ZHOU De-shan
Department of Histology and Embryology, College of Medicine, Third Military Medical University, Chongqing 400038, China
关键词:
单克隆抗体黏蛋白结直肠癌
Keywords:
monoclonal antibody mucin colorectal carcinoma
分类号:
R392-33;R392.2;R735
文献标志码:
B
摘要:
目的    制备结直肠癌特异性单克隆抗体,为深入研究相关肿瘤的发病机制提供新的实验工具。    方法    用人结直肠癌患者的手术标本制备细胞悬液免疫小鼠,按常规方法行细胞融合,经免疫组织化学染色筛选阳性克隆,并对其抗体的特性和生物学性状等进行分析。    结果    共获得32个阳性克隆,其中针对结直肠癌较特异性的阳性克隆有2个(克隆株2-3,15-8)。免疫组化染色显示,克隆株2-3培养上清和腹水与结直肠癌组织的冰冻切片反应的阳性率可达100%。癌旁正常组织的阳性率较低,且染色强度远弱于癌组织,当抗体稀释度超过1∶6 000时反应阴性,而肿瘤组织仍呈强阳性。从结肠癌的分化程度来看,分化较差的肿瘤组织,包括中、低分化的癌组织染色较强。另外,免疫染色阳性产物主要位于肿瘤的腺腔和正常黏膜表面,提示该抗体所识别的抗原可能为结直肠癌细胞和黏膜上皮分泌的黏蛋白等抗原。    结论    本研究成功地制备了两株针对结直肠癌的较特异的单克隆抗体,可作为实验研究相关蛋白在结直肠癌及其他肿瘤发病中的作用机制的方法。
Abstract:
Objective    To produce monoclonal antibodies against colorectal carcinoma and provide a new tool for study of the mechanisms in related cancers.     Methods    Specimens of colorectal carcinoma were freshly obtained from surgery. Balb/c mice were immunized with cell suspension from colorectal carcinoma tissues. Fusion was done as classical hybridoma method. Positive hybridomas were selected by immunohistochemical staining. The nature and biological characteristic of the selected monoclonal antibodies was investigated.     Results    Thirty-two positive hybridomas were selected. Two (2-3,15-8) of these hybridomas secreted antibodies binding specifically to human colorectal carcinoma. By immunohistochemical staining, all of colorectal carcinoma cases was positive for 2-3 antibody. In contrast, normal colon and rectal mucosa showed much weaker staining, and was totally negative when 2-3 ascites antibody was diluted over 1∶6 000. In addition, 2-3 antibody reacted strongly with moderately differentiated and poorly differentiated colorectal carcinoma. The immunoreactive substance was observed on luminal contents of colorectal carcinoma tubules and on apical surface of normal epithelium, indicating that the antigen recognized by 2-3 antibody may be a kind of mucin secreted by colorectal carcinoma and normal epithelium and may participate in tumor invasion and metastasis.     Conclusion    We successfully produced two monoclonal antibodies against colorectal carcinoma.

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更新日期/Last Update: 2008-10-24