中国临床康复2004，Vol.8Issue(33)：7606-7608,3.
应用细胞工程和转基因技术治疗糖尿病:"胰岛代理细胞"的构建研究
Construction of agent islet cell with cell engineering and transgeneic technology on the treatment of diabetes mellitus
摘要
Abstract
BACKGROUND: At present, good cell source for transplant treatment of diabetes is limited, because it is hard to get β cell.OBJECTIVE: To construct"artificial beta cells" which exhibit physiologic glucose-stimulated insulin secretion for effective cure of diabetes.DESIGN: A Randomly controlled trial.SETTING, MATERIAL and INTERVENTIONS: This study was conducted in the center lab of the Third Military Medical University Affiliated Xinqiao Hospital. HepG2 cells were infected with recombinant retrovirus carrying glucokinase(GK)gene and mutant proinsulin(MPIN)gene, then selectively cultured with G418 and identified by radioimmunoassay,SDS-PAGE, Western-blotting and RT-PCR techniques. At last, the dose-response effect of glucose on insulin secretion from HepG2 cells expressed both GK and MPIN gene was tested with HepG2 cells that only expressed MPIN as controls.MAIN OUTCOME MEARSURES: The concentration of insulin was measured in cultivation supernatant of combined expression HepG2 at different glucose levels.RESULTS: HepG2 cells with GK gene and MPIN gene transferred were selectively cultured with G418 and the positive clones obtained in 3 weeks. GK gene and MPIN gene integration and expression were detected by radioimmunoassay, SDS-PAGE, Western-blotting and RT-PCR techniques. The "artificial beta cell" obtained a glucose-stimulated insulin secretion with maximal insulin secretion at 1.75-2.00 mmol/L glucose concentration. While the insulin secretion in HepG2 cell with MPIN gene expression was lack of glucose responsiveness( P ＞ 0.05).CONCLUSION: An artificial beta cell was successfully constructed.关键词
基因方法/糖尿病/治疗/逆转录病毒科/胰岛/细胞学分类
医药卫生引用本文复制引用
郑宏庭,邓华聪,蹇锐,兰丽珍,方芳..应用细胞工程和转基因技术治疗糖尿病:"胰岛代理细胞"的构建研究[J].中国临床康复,2004,8(33):7606-7608,3.基金项目
国家自然科学基金资助项目(30070356) （30070356）
