pIRES2-FL-IL-3转染协同促进脐血干细胞增殖的研究  

pIRES2-FL-IL-3 transfection promotes proliferation of cord blood stem cells

作　　者：张勇[1] 张林生[1] 郭朝华[2] 张红宾[1] 董世武[2] 

机构地区：[1]第三军医大学,西南医院血液科 [2]第三军医大学军事预防医学院防原医学教研室,全军复合伤研究所,创伤,烧伤与复合伤国家重点实验室,重庆400038

出　　处：《第三军医大学学报》2007年第24期2330-2332,共3页Journal of Third Military Medical University

基　　金：国家自然科学基金青年基金(30200071)~~

摘　　要：目的观察真核共表达质粒载体pIRES2-FL-IL-3转染对脐血干细胞增殖的影响及目的蛋白IL-3和FL(Flt3ligand)的表达变化。方法分为4组:空白对照组(对照组)、pIRES2-IL-3组(IL-3组)、pIRES2-FL组(FL组)和pIRES2-FL-IL-3组(共表达组)。通过台盼蓝染色及MTT法检测pIRES2-FL-IL-3转染(共表达组)对脐血干细胞细胞存活率及增殖的影响,RT-PCR技术、Western blot技术检测脐血干细胞FL和IL-3的mRNA水平和蛋白表达。结果共表达组细胞存活率无显著变化(P>0.05),MTT法检测细胞增殖显著高于对照组,共表达组IL-3及FL mRNA水平和蛋白均显著高于对照组,并且高于IL-3组和FL组。真核共表达质粒载体pIRES2-FL-IL-3转染脐血干细胞后细胞存活率无显著变化(P>0.05),结论脐血干细胞增殖显著增强,目的蛋白能得到有效表达。Objective To observe the proliferation of cord blood stem cells and detect their mRNA and protein levels of FL and IL-3 after transfection of plasmid vector pIRES2-FL-IL-3. Methods The cell survival rate and proliferation of the cord blood stem cells were observed by dye staining and MTT methods after transfected by plasmid vector pIRES2-IL-3, pIRES2-FL or pIRES2-FL-IL-3. The mRNA and protein levels of FL and IL-3 were detected by RT-PCR and Western blot. Results No statistical difference on cell survival rate of cord blood stem cells was observed between those transfected by plasmid vector pIRES2-FL-IL-3 and their corresponding controls. The cell proliferation of cord blood stem cells transfected by pIRES2-FL-IL-3 was higher than those transfected by pIRES2-IL-3 or pIRES2-FL. The mRNA and protein levels of FL and IL-3 in the cord blood stem cells transfected by pIRES2-FL-IL-3 were higher than those in the normal cord blood stem cells, and those transfected pIRES2-IL-3 group or pIRES2-FL respectively. Conclusion The transfection of pIRES2-FL- IL-3 to the cord blood stem cells does not influence the cell survival rate but accelerates their proliferation. FL and IL-3 co-expression displays synergic effect on the proliferation of cord blood stem cells.

关 键 词：白介素3 FLT3配体 真核表达 增殖 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R392.11[医药卫生—基础医学]