含有RU486调控系统的质粒DNA介导的可诱导的IL-12基因在小鼠体内表达  

Inducible IL-12 gene expression mediated by naked DNA containing an RU486 regulatory system

作　　者：李艳茹[1] 张海英[1] 王琳[1] 李晓敏[1] 王健君[1] 李玉林[1] 

机构地区：[1]吉林大学基础医学院病理学教研室 病理生物学教育部重点实验室,长春130021

出　　处：《中华微生物学和免疫学杂志》2008年第2期102-106,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金资助项目（30370547）;教育部新世纪优秀人才支持计划资助（2004）;教育部优秀青年教师资助计划项目（2003）

摘　　要：目的评价含有RU486调控系统的质粒DNA诱导目的基因表达的能力。方法将编码LacZ的pRS-LacZ和编码小鼠IL-12的pRS22体外转染细胞或静脉快速注射入小鼠体内。X-gal染色检测培养细胞和组织中LacZ基因的表达。ELISA法检测培养细胞上清和血清中IL-12的表达水平。结果加入RU486诱导后，在肝脏来源的HepG2细胞中约30％被染成蓝色；而不加RU486时．仅有不到1％的细胞蓝染。在其他非肝脏来源的细胞中，无论是否加入RU486诱导，都只有不到0．5％的细胞蓝染。转染pRS22的HepG2细胞，培养细胞上清中IL-12表达水平随RU486浓度的升高而增加。静脉注射pRS-LacZ后，用RU486诱导时，仅在肝脏组织中见到蓝染细胞。静脉注射pRS22后，血清中IL-12表达水平与质粒及RU486的剂量有关。结论含有RU486调节系统的质粒DNA可以通过加入或取消RU486来开启或关闭IL-12基因的表达，而且可以通过改变质粒或RU486的剂量来调控IL-12基因的表达水平。Objective To evaluate the inducibility of target gene expression induced by plasmid DNA carrying a RU486 regulatory system. Methods Plasmid pRS-LacZ encoding LacZ gene and plasmid pRS22 encoding murine interleukin-12 （IL-12） gene were used to transfect cells in vitro or be rapidly injected into the vein of mice. Then LacZ expression was assayed in cultured cells and tissues by X-gal staining. The IL-12 level was tested in the supernatants of cultured cells and in the serum with ELISA. Results In the presence of RU486, 30% of the liver derived hepatic HepG2 cells were dyed blue while less than 1% of the HepG2 cells were blue colored in the absence of RU486. Less than 0.5% of other non-hepatic cells were in blue with or without RU486. After transfection, the IL-12 level in the supernatants of HepG2 cells increased with increasing concentrations of RU486 treatment. After intravenous injection of plasmid pRS-LacZ, blue colored cells were observed only in liver tissue while induced by RU486. After intravenous injection of plasmid pRS22, the IL-12 expression level in serum was related with the dosage of RU486 or/and plasmid DNA. Conclusion Plasmid DNA containing RU486 regulatory system could open or close IL-12 gene expression by the addition or deletion of RU486. And the IL-12 expression level in serum could be regulated by altering the dose of plasmid DNA or/and RU486.

关 键 词：基因调节 RU486 质粒DNA 高压注射 

分 类 号：R686[医药卫生—骨科学]