重组腺病毒介导的白细胞介素-12(IL-12)在肝癌细胞中的表达  

ADENOVIRUS MEDIATED EXPRESSION OF INTERLEUKIN-12 IN HEPATOMA CELLS

作　　者：王自力[1] 梁国栋[1] 付士红[1] 周国林[1] 刘礼初 陈飞[1] 施侣元[1] 侯云德[1] 

机构地区：[1]中国预防医学科学院病毒学研究所病毒基因工程国家重点实验室

出　　处：《病毒学报》1999年第2期119-124,共6页Chinese Journal of Virology

摘　　要：利用腺病毒表达系统在肝癌细胞中成功地表达了有生物活性的白细胞介素－１２（ＩＬ－１２）。ＩＬ－１２的Ｐ３５和Ｐ４０ｃＤＮＡ分别克隆到腺病毒载体ｐＡＣＣＭＶｐＬｐＡ，构建ｐＡＣ／Ｐ３５和ｐＡＣ／Ｐ４０表达质粒。与腺病毒重组质粒ｐＪＭ１７共转染２９３细胞，通过基因重组产生ＩＬ－１２Ｐ３５和Ｐ４０重组腺病毒。用重组腺病毒感染肝癌细胞株ＨｅｐＧ２和ＳＭＭＣ７７２１，经ＥＬＩＳＡ和Ｗｅｓｔｅｒｎ检测证明，在这两株细胞中均有ＩＬ－１２的表达，并且经ＩＦＮ－γ诱生法检测表明，在ＨｅｐＧ２和ＳＭＭＣ７７２１细胞中表达的ＩＬ－１２具有诱导产生ＩＦＮ－γ的生物学活性。本实验为研究ＩＬ－１２对肝癌的基因治疗奠定了基础。This experiment was designed to study the adenovirus mediated expression of interleukin-12 (IL-12) in hepatoma cells. The two subunits of IL-12, P35 and P40, were cloned into adenovirus vector pACCMV. pLpA by using the adenovirus expression system. The recombinant expression plasmids of P35 and P40, pAC/P35 and pAC/P40, were constructed respectively. The two plasmids were cotransfected into 293 cell line with adenovirus gene recombinant plasmids pJM17. The IL-12 P35 and P40 recombinant adenovirus were produced by gene recombination. Expression of IL-12 was proved in the HepG2 and SMMC7721 cells by ELISA and Western blot. IFN-γ induction test indicated that IL-12 expressed in two cells had the biological activity of IFN-γ induction. By using adenovirus expression system, IL-12 with biological activity was expressed successfully in hepatoma cells. This experiment laid a foundation for gene therapy of hepatoma with IL-12.

关 键 词：白细胞介素12 基因治疗 基因表达 肝癌 腺病毒 

分 类 号：R392.11[医药卫生—免疫学] R730.2[医药卫生—基础医学]