重组人IL-24的表达及其体外抗肿瘤细胞活性的研究  

Expression of recombinant human interleukin-24 and its anti-tumor activity in vitro

作　　者：刘朝阳[1] 瞿爱东[1] 徐帆洪[1] 

机构地区：[1]上海生物制品研究所第一研究室,200052

出　　处：《国际生物制品学杂志》2009年第4期175-179,共5页International Journal of Biologicals

摘　　要：目的构建重组人白细胞介素24（recombinant human interleukin24，rhIL-24）表达载体并在大肠杆菌中表达，体外评估rhIL-24的生物学活性。方法用基因工程方法构建rhIL-24表达载体并在大肠杆菌中表达rhIL-24。表达的重组蛋白经层析法纯化后，分别用噻唑蓝比色法和蛋白印迹法检测rhIL-24对肿瘤细胞生长的影响和对内源性IL-24的诱导，用实时PCR检测不同肿瘤细胞与rhIL-24共孵育后细胞内的胱天蛋白酶3（caspase3）mRNA变化。结果rhIL-24能在大肠杆菌中高效表达，并能明显抑制多种肿瘤细胞生长。rhIL-24能诱导正常MRC-5细胞和多种肿瘤细胞产生内源性IL-24，并能改变肿瘤细胞的胱天蛋白酶3水平。结论rhIL-24能在大肠杆菌中高效表达，且具有生物学活性。Objective To construct an expression vector of recombinant human interleukin-24 （rhIL-24） and express it in E. coli, and evaluate the biological activity in vitro. Methods An expression vector of rhIL-24 was constructed by genetic engineering and rhIL-24 was expressed in E. coll. After purification of rhIL-24 by chromatography, MTT assay was used to determine the effect of rhIL-24 on the growth of tumor cells and the Western blotting was used to determine the induction of endogenous IL-24 by rhIL-24, rhIL-24 was coincubated with different tumor cells, and real-time PCR was used to determine the change of easpase 3 mRNA. Results rhIL-24 was highly expressed in E. coli, and the growth of tumor cells was inhibited significantly by rhIL-24, rhIL-24 could induce normal MRC-5 ceils and tumor cells to produce endogenous IL-24, and change caspase 3 level of tumor cells. Conclusion rhIL-24 is highly expressed in E. coli and has biological activity.

关 键 词：白细胞介素类 抗肿瘤药 细胞凋亡 半胱氨酸天冬氨酸蛋白酶3 

分 类 号：R9[医药卫生—药学]