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。
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 eu- karyotic 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 eu- karyotic expression vector is successfully constructed which can be an important tool for the further study.
出处
《山东大学学报（医学版）》
CAS
北大核心
2012年第12期19-24,共6页
Journal of Shandong University:Health Sciences
基金
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)
关键词
肝癌
免疫组化
A20真核表达载体
NF-ΚB信号通路
Human hepatocellular carcinoma
Immunohistochemistry
A20 eukaryotic expression vector
NF-κB signal pathway
