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山东大学学报(医学版) ›› 2012, Vol. 50 ›› Issue (12): 19-.

• 基础医学 • 上一篇    下一篇

免疫负调控基因A20在肝癌中的表达及其真核表达载体的构建

邢巧玲1, 韩振龙2, 宋永红3, 郭春1, 朱法良1, 张利宁1, 石永玉1   

  1. 1. 山东大学医学院医学免疫学研究所, 济南 250012;
     2.山东省警官总医院检验科,  济南 250002;    3. 山东省血液中心, 济南 250014
  • 收稿日期:2012-07-18 出版日期:2012-12-10 发布日期:2012-12-25
  • 通讯作者: 石永玉(1970- ), 男, 副教授, 主要从事肿瘤免疫的研究。E-mail:shiyongyu@sdu.edu.cn
  • 作者简介:邢巧玲(1983- ), 女, 硕士研究生, 主要从事肝癌免疫机制的研究。E-mail:xing-qiao-ling@126.com
  • 基金资助:

    国家自然科学基金 (81071705);山东省中青年科学家科研奖励基金 (BS2009YY002)

Expression of negative immunoregulatory gene A20 in human
hepatocellular carcinoma  and construction of
its eukaryotic expression vector

XING Qiao-ling1,   HAN Zhen-long2, SONG Yong-hong3, GUO Chun1,
ZHU Fa-liang1, ZHANG Li-ning1, SHI Yong-yu1   

  1. 1. Institute of Immunology, School of Medicine, Shandong University,  Jinan 250012,   China;
    2.Clinical laboratory of Shandong Police Hospital,  Jinan 250002,  China;
    3. Blood Center of Shandong Province, Jinan 250014,  China
  • Received:2012-07-18 Online:2012-12-10 Published:2012-12-25

摘要:

目的    探讨免疫负性调控基因A20在肝癌中的表达并构建A20真核表达载体。 方法   采用免疫组化技术, 检测46例肝癌患者癌组织和癌旁组织中A20的表达情况, 并进一步构建A20真核表达载体, 测序正确后以脂质体法转染至SMMC-7721肝癌细胞系中,  RT-PCR和Western blot检测转染后A20的mRNA和蛋白表达。结果   免疫组化结果显示, A20在肝癌组织中的表达明显低于癌旁组织, 差异具有统计学意义(P<0.05)。A20的表达与肝癌分级分化和肿瘤大小有关(P<0.05), 成功构建A20真核表达载体, 并在SMMC7721肝癌细胞系中获得高表达。结论   免疫负调控基因A20可能参与了肝癌的发生机制, 成功构建的A20真核表达载体可作为进一步研究的重要工具。

关键词: 肝癌;免疫组化;A20真核表达载体;NF-κB信号通路

Abstract:

Objective   To explore the role of negative immunoregulatory gene A20 in human hepatocellular carcinoma(HCC), we investigated the expression of A20 gene in HCC and constructed the A20 eukaryotic expression vector. Methods   The expressions of A20 gene in tumor tissues and tumor-adjacent tissues from 46 HCC patients were detected by immunohistochemistry. The A20 eukaryotic expression vector was constructed and transfected into SMMC-7721 HCC cell lines by the liposome. The mRNA and the protein levels of A20 were detected by the RT-PCR and Western blot methods. Results   A20 expression in tumor tissues was significantly lower than that in tumor-adjacent tissues(P<0.05). A20 expression was closely associated with the tumor differentiation degree and size (P<0.05). The A20 eukaryotic expression vector was successfully constructed which induced the overexpression of A20 in SMMC -7721 cell lines. Conclusion   Negative immunoregulatory gene A20 may be involved in the tumorigenesis of HCC. The A20 eukaryotic expression vector is successfully constructed which can be an important tool for the further study.

Key words: Human hepatocellular carcinoma; Immunohistochemistry; A20 eukaryotic expression vector; NF-κB signal pathway

中图分类号: 

  • R392.3
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