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预防医学  2019, Vol. 31 Issue (3): 231-235    DOI: 10.19485/j.cnki.issn2096-5087.2019.03.004
  论著 本期目录 | 过刊浏览 | 高级检索 |
ANKRD22检测抗体制备及其在结直肠癌中的表达
林云华1, 柳景文2, 杨赛赛2, 刘希永3, 王宏平2
1.平湖市第一人民医院,浙江 平湖 314200;
2.浙江大学医学院附属第二医院;
3.杭州贺普生物有限公司
Preparation of antibody against ANKRD22 and its expression in colorectal cancer
LIN Yun-hua*, LIU Jing-wen, YANG Sai-sai, LIU Xi-yong, WANG Hong-ping
*First People's Hospital of Pinghu,Pinghu,Zhejiang 314200,China
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摘要 目的 制备特异性锚蛋白重复结构域蛋白22(ANKRD22)抗体,了解ANKRD22在结直肠癌中的表达。方法 用pET-42a表达系统从大肠杆菌中表达重组人ANKRD22蛋白,纯化后免疫Balb/c小鼠,以高表达ANKRD22的293T细胞裂解液为抗原,采用免疫印迹法筛选抗人ANKRD22特异性单克隆抗体;采用免疫组织化学染色法检测112例结直肠癌患者组织芯片中ANKRD22的表达。结果 本研究获得了93个杂交瘤细胞,筛选出4株抗人ANKRD22特异性单克隆抗体,均与天然的人ANKRD22呈良好反应。ANKRD22主要分布于结直肠癌细胞的细胞质。在112例患者结直肠癌组织中,ANKRD22表达阳性94例,阳性率为83.93%。ANKRD22表达阳性与p53表达和β-catenin表达存在统计学关联(P<0.05),而与年龄、性别、结直肠肿瘤位置、临床AJCC分期、Dukes分期、肿瘤分化程度、有无淋巴结转移和错配修复基因表达均无统计学关联(P>0.05)。结论 ANKRD22在结直肠癌组织中表达水平较高,可能参与了结直肠上皮的癌变过程,是一个潜在的辅助诊断标志。
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林云华
柳景文
杨赛赛
刘希永
王宏平
关键词 结直肠癌ANKRD22表达单克隆抗体免疫组织化学染色法    
AbstractObjective To prepare the monoclonal antibody against ankyrin repeat domain 22(ANKRD22)and to investigate its expression in colorectal cancer tissues.Methods The recombinant human ANKRD22 was expressed through E. coli and pET-42a and then used to immunize Balb/c mice after purification. Anti-human ANKRD22 specific monoclonal antibodies were selected by Western blotting with 293T cell lysate highly expressing ANKRD22 as antigen. The expression of ANKRD22 in the tissue microarrays of 112 patients with colorectal cancer was detected by immunohistochemical staining.Results Four specific monoclonal antibodies against human ANKRD22 were screened out of 93 hybridoma cells,which reacted well with natural human ANKRD22. ANKRD22 was mainly distributed in the cytoplasm of colorectal cancer cells. In 112 cases of colorectal cancer,94 cases were detected positive for ANKRD22 expression,with the positive rate of 83.93%. The expression of ANKRD22 was statistically correlated with the expression of p53 and β-catenin(P<0.05),but not with age,sex,location of tumors,AJCC stage,Dukes stage,degree of differentiation,lymph node metastasis and mismatch repair gene expression(P>0.05).Conclusion The expression level of ANKRD22 was high in colorectal cancer. ANKRD22 might be involved in the carcinogenesis of colorectal epithelium and be a potential diagnostic marker.
Key wordsColorectal cancer    ANKRD22    Expression    Monoclonal antibody    Immunohistochemical staining
收稿日期: 2018-11-14      修回日期: 2018-12-30      出版日期: 2019-03-04
中图分类号:  R735.3  
作者简介: 林云华,本科,副主任技师,主要从事医学检验工作
通信作者: 王宏平,E-mail:wanghp02@163.com   
引用本文:   
林云华, 柳景文, 杨赛赛, 刘希永, 王宏平. ANKRD22检测抗体制备及其在结直肠癌中的表达[J]. 预防医学, 2019, 31(3): 231-235.
LIN Yun-hua, LIU Jing-wen, YANG Sai-sai, LIU Xi-yong, WANG Hong-ping. Preparation of antibody against ANKRD22 and its expression in colorectal cancer. Preventive Medicine, 2019, 31(3): 231-235.
链接本文:  
https://www.zjyfyxzz.com/CN/10.19485/j.cnki.issn2096-5087.2019.03.004      或      https://www.zjyfyxzz.com/CN/Y2019/V31/I3/231
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