摘要
目的:构建重组真核表达质粒pIRESEgr-IFNγ并检测其在体外培养的Lewis肺癌细胞中的辐射诱导表达。方法:利用基因重组技术构建含Egr-1启动子的IFNγ表达质粒,脂质体介导体外转染Lewis肺癌细胞,酶联免疫吸附法(ELISA)检测0、2、5和10Gy X射线诱导IFNγ表达的量效和时程关系。结果:酶切鉴定证实,Egr-1启动子和IFNγ基因正确插入真核表达载体pIRES1neo;2、5和10Gy X射线照射转染该重组质粒的Lewis肺癌细胞,上清中IFNγ表达量明显高于假照组(P<0.001),其中5Gy照射后表达量最高,为假照组的4.39倍。5Gy照射后2、6、12和36h上清中IFNγ表达量逐渐增加(P<0.001),照射后36h表达量为照射前的6.27倍。结论:成功构建了真核表达质粒pIRESEgr-IFNγ,该质粒具有辐射诱导基因表达增强特性。
Objective To construct the recombinant plasmid plRESEgr-IFNγ and detect its expression in Lewis lung carcinoma induced by irradiation in vitro. Methods The recombinant plasmid pIRESEgr-IFNγ containing Egr-1 promoter and IFNγ gene was constructed with gene recombinant technique. The plasmid was transferred into Lewis lung carcinoma by liposome in vitro. The correlations of dose- and time-effects in the expression of IFNγ gene induced by X-ray were detected by ELISA. Results The identification with enzymes proved that Egr-1 promoter and IFNγ gene were inserted into vector pIRESlneo correctly. After X-ray irradiation with different doses, the expression of IFNγ in the supernatant of Lewis lung carcinoma transfected by pIRESEgr-IFNγ was significantly higher than that in 0 Gy group （P〈0. 001）. After 5 Gy X-ray irradiation, the expression of IFNγ was the highest, being 4.39 times as much as that in 0 Gy group. The expression of IFNγ in the supernatant increased after 5 Gy X-ray irradiation, being 6.27 times as much as that in 0 h group 36 h after irradiation. Conclusion The recombinant plasmid pIRESEgr-IFNγ is constructed successfully, and it has the property of enhancing the expression of IFNγ gene induced by irradiation.
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2007年第5期790-793,共4页
Journal of Jilin University：Medicine Edition
基金
国家自然科学基金资助课题(30600160
30570546)
