人皮肤成纤维细胞体外转染Brgl基因的实验研究

目的探讨重组Brgl基因转染人皮肤成纤维细胞的可行性，以及转染对细胞增殖和活性的影响。方法体外重组Brgl基因，借助真核表达载体系统，转入体外培养的人皮肤成纤维细胞；通过流式细胞仪检测报告基因表达，并确定细胞转染效率：应用ReahimePCR比较转染前后BrglmRNA的表达；MTT法检测Brgl基因对细胞增殖能力的影响。结果Brgl基因转染后，（73．0±6．7）％的被转染细胞表达报告基因；BrglmRNA在转染组、转染空载体组、未转染组细胞中的表达分别为（6．23±1．18）、（1．11±0．22）和（1．52±0．12），转染组BrglmRNA相对表达量较未转染组及转染空载体组明显增高（P〈0．01）；细胞的增殖能力在Brgl基因转染前后无显著差异。结论人皮肤成纤维细胞可作为Brgl转染的靶细胞．转染Brgl基因对人成纤维细胞的增殖能力无显著影响。

Transfection of Recombinant Brgl Gene into Human Skin Fibroblasts in Vitro

Objective To explore the feasibility of transfecting recombinant Brgl into human skin fibroblasts and to investigate the impact on cell proliferation rate and activity. Methods Recombinant human Brgl was transfected into skin fibroblasts by the karyocyte expressive vector. The cell transfection efficiency was determined through testing reported gene expression by flow eytometry; The expression of Brgl mRNA before and after transfection was quantified by Reahime PCR; MTT was used to detect cell proliferation before and after transfection. Results The cell transfection efficiency of human skin fibroblasts was nearly （73.0±6.7）%; The relative expression level of Brgl mRNA in transfected cell group, empty-vector cell group or un-transfected cell group were respectively 6.23 ±1.18, 1.11±0.22 and 1.52±0.12. The level of mRNA in transfected cell group was significantly higher compared with either un-transfected cell group or empty-vector cell group （P〈0.01）; The proliferation abilities had no significant difference before and after transfection. Conclusion The human skin fibroblasts can be used as the target cells of Brgl gene transfection, which has no significant impact on fibroblast proliferation.