【摘要】 目的构建双基因表达质粒pEgr-IFNγ-endostatin并检测其在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和endostatin双基因表达质粒,脂质体介导体外转染Lewis肺癌细胞,用酶联免疫吸附法(ELISA)检测不同剂量X射线诱导IFNγ和endostatin表达的量效关系和时程。结果酶切鉴定证实Eg-r1启动子、IFNγ和endostatin基因正确插入双基因表达载体pIRES1neo;不同剂量X射线照射转染该重组质粒的Lewis肺癌细胞上清中IFNγ和endostatin表达量明显高于假照组(均P<0.001),其中5Gy照后表达量最高,分别为假照组的4.14倍和2.92倍;2GyX射线照后Lewis肺癌细胞上清中IFNγ和endostatin含量随时间延长而增加,照后36h分别为照射前的3.75倍和3.02倍(均P<0.001)。结论本实验成功构建了双基因表达更多还原

【Abstract】 Objective To construct a recombinant dual_gene co_expressing plasmid pEgr_IFN γ_endostatin and detect its radiation_induced expression in Lewis lung cancer cells. Methods The recombinant plasmid pEgr_IFN γ_endostation containing Egr₁ promoter,IFNγ and endostatin genes was constructed with gene recombination technique.The plasmid was transferred into Lewis lung cancer cells by liposme in vitro. The correlation of dose and effects and the time_course patterns of the expressions of IFNγ and endostatin genes induced by different doses of X_rays were detected by ELISA. Results Identification with enzymes proved that Egr₁ promoter,IFNγ and endostatin genes were inserted into the dual_gene co_expressing vector pIRESlneo correctly.After different doses of X_irradiation,the expressions of IFNγ and endostatin the supernatant of cultured Lewis lung cancer cells transfected by pEgr_IFNγ_endostatin were significantly higher than those in 0 Gy group.After 5 Gy X_irradiation,the expressions of IFNγ and endostatin were the highest,being 4^14 and 2^92 times as much as those in 0 Gy group respectively.The concentrations of IFNγ and endostatin in the supernatant increased after 2 Gy X_irradiation,being 3^75 and 3^02 times as much as those in 0 Gy group respectively 36 h after irradiation （P<0^001）. Conclusion The dual_gene co_expressing plasmid pEgr_IFN γ-endostatin has been constructed successfully,and it has the property of enhancing the co_expression of IFNγ and endostatin genes induced by irradiation.;更多还原