摘要
[目的]检测从人白介素-12(human interleukin 12,hIL-12)转基因马铃薯中提取的hIL-12的抗肝癌效应。[方法]用MTT法检测转基因马铃薯中hIL-12对人肝癌细胞HepG22.2.15的体外杀伤作用;用AnnexinV/PI双染色法加激光共聚焦显微镜观察转基因马铃薯中hIL-12对HepG22.2.15细胞的诱导凋亡作用。[结果]转基因马铃薯中表达的hIL-12与重组人白介素-12(rhIL-12)在体外对HepG22.2.15具有杀伤作用,而野生对照组无杀伤作用;An-nexinV/PI双染色法加激光共聚焦显微镜观察结果表明,转基因马铃薯中表达hIL-12可诱导HepG22.2.15细胞凋亡。[结论]转基因马铃薯表达体系表达的人白介素-12具有抗肝癌细胞增殖、促进肝癌细胞凋亡的作用。
[Objective]The aim was to examine the anti heptocarcinoma effects of human interleukin-12（hIL-12） from transgenic potatoes.[Method]Human heptocarcinoma cell line HepG22.2.15 was cocultured with human peripherial blood monocyte（PBMC）.Human IL-12 extracted from its transgenic patatoes was introduced to this coculture system.MTT method and laser confocal microscope were then employed to evaluate its survival rates and morphological changes of HepG22.2.15 cell line.[Result]The HepG22.2.15 survival rate of the plant produced hIL-12 treated group was significantly lower than that of the wild type control,while similar to that of commercial purified recombinant hIL-12 group.As indicated by AnnexinV/PI double staining laser confocal microscope,cocultured heptocarcinoma cells showed the typical early and final phase apoptotic morphological characteristics 48 hours after 2 × 10-4 mg/L potato secreted hIL-12 treatment.[Conclusion] These results demonstrated that Heptocarcinoma HepG22.2.15 cell growth could be actively inhibited by the transgenic potato expressed hIL-12 in vitro.
基金
Supported by the Key Project of Education Department of Guizhou Province "Establishment of Human Interleukin-12 Plant Expression System" (2004118)~~
