sTRAIL真核表达载体的构建及其对裸鼠胶质瘤的治疗作用

Construction of eukaryotic recombinant sTRAIL expression vector and its effect on C6 glioma xenograft in athymic mice

目的构建重组基因表达载体PcDNA-sTRAIL并观察其对裸鼠种植性胶质瘤生长的抑制作用。方法通过PCR扩增和定向克隆技术构建重组真核表达载体PcDNA-sTRAIL。以阳离子脂质体介导转染裸鼠种植性胶质瘤细胞,同时设立未转染、空质粒转染及Lipofectin转染组,检测转染阳性率,并观察该基因载体对种植瘤生长的抑制作用。结果经过测序鉴定和蛋白表达检测证实,重组真核表达载体PcDNA-sTRAIL构建成功。阳离子脂质体介导质粒转染种植瘤瘤细胞阳性率达50%。脂质体PcDNA-sTRAIL转染种植瘤细胞后,相比未转染、空质粒转染及Lipofectin转染组,种植瘤体积、重量增长明显受抑制(均P<0.05),表现出一定的肿瘤细胞增殖抑制作用。结论本研究成功构建了sTRAIL基因的真核表达载体,并从体外实验中初步证实了其对裸鼠种植性胶质瘤细胞增殖具有明显的抑制作用。

Objective To construct eukaryotic recombinant sTRAIL expression vector and to investigate its effect on C6 glioma xenograft in athymic mice. Methods Eukaryotic recombinant sTRAIL expression vector PcDNA-sTRAIL was constructed by means of PCR and directed cloning. PcDNA-sTRAtL was transfected to C6 glioma cells mediated by cationic liposome and the positive transfection rate was detected by immunohistochemistry. C6 glioma xenografts in athymic mice were treated with PcDNA-sTRAIL plus lipofectin or blank vector and lipofectin as control groups. The weight and volume of xenografts in each groups were compared. Results The construction of eukaryotic recombinant sTRAtL expression vector PcDNA-sTRAIL was confirmed by gene sequencing and Western blot. After transfection of PcDNA-sTRAIL, up to 50 % of C6 cells positively expressed sTRAIL protein. In the in vivo study, both weight and volume in PcDNA-sTRAIL plus Lipofectin group were significantly lower than those in control groups （P〈0.01）. Conclusion A eukaryotic recombinant sTRAIL expression vector was successfully constructed and it can inhibit the growth of C6 glioma xenograft in athymic mice.