hIGF-1基因转染对成纤维细胞增殖的影响

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。

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

Objective 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 G_1 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.