hIGF-1基因转染对成纤维细胞增殖的影响  

Effects of human insulin-like growth factor 1 gene transfection on proliferation of NIH3T3 fibroblasts

作　　者：张绍昆[1] 谭岩[2] 单玉兴[1] 宋之明[1] 徐莘香[1] 

机构地区：[1]吉林大学第一医院骨科,吉林长春130021 [2]吉林大学第一医院中心实验室,吉林长春130021

出　　处：《吉林大学学报（医学版）》2006年第2期210-213,共4页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金资助课题(30400447)

摘　　要：目的:探讨人胰岛素样生长因子1(hIGF-1)基因转染对成纤维细胞增殖的影响。方法:用脂质体转染法将pcDNA3.1-hIGF-1质粒转染鼠成纤维细胞NIH3T3,经G418筛选抗性NIH3T3细胞并继续培养4周,进行原位杂交和免疫细胞化学检测hIGF-1的表达,MTT方法和流式细胞仪检测NIH3T3细胞增殖能力。结果:转染pcDNA3.1-hIGF-1后的NIH3T3细胞内有大量hIGF-1 mRNA和蛋白质的表达;MTT检测显示转染pcDNA3.1-hIGF-1的NIH3T3细胞光吸收值增大,与未转染的NIH3T3细胞组比较,差异具有显著性(P<0.01);流式细胞仪检测显示转染组细胞,S期比例增加(59.3%),G1期比例减少(27.2%)。结论:pcDNA3.1-hIGF-1质粒转染成纤维细胞后,可以获得稳定表达,并能明显促进成纤维细胞的增殖。Obioctive To study the effects of human insulin-like growth factor 1 （hIGF-1） gene transfection on the proliferation of NIH3T3 fibroblasts. Methods The plasmid of pcDNA3.1-hIGF-1 was transfected into NIH3T3 fibroblasts by using Lipofectin method. The positive cell clones were selected with G418 and cultured for 4 weeks. The stable expression of hIGF-1 in the positive cells was determined by in situ hybridization and immunocytochemical analysis. MTT assay and flow cytometer analysis were used to observe the proliferation of NIH3T3 fibroblasts. Results hIGF-1 mRNA and protein expressed in NIH3T3 fibroblasts transfected with pcDNA3.1-hIGF-1 by in situ hybridization and immunocytochemical analysis. MTT assay showed the A value of transfected NIH3T3 fibroblasts rose, compared with untransfected NIH3T3 fibroblasts group, the difference was significant （P〈0.01）. Cellular proportion in S period （59.3%） was increased and it in G1 periods was decreased （27.2%） after transfection by flow cytometer measurement. Conclusion The stable expression of hIGF-1 in NIH3T3 fibroblasts transfected with pcDNA3.1-hIGF-1 is obtained. Gene transfection of hIGF-1 can stimulate the proliferation of NIH3T3 fibroblasts.

关 键 词：胰岛素样生长因子1 基因表达 成纤维细胞 3T3细胞 细胞周期 

分 类 号：Q78[生物学—分子生物学]