小干扰RNA分子对增生性瘢痕成纤维细胞增殖的影响

目的 使用RNA干扰（RNA interference，RNAi）技术抑制黏着斑激酶（Focal adhesion kinase，FAK）基因的表达，观察增生性瘢痕成纤维细胞在生物学行为上发生的变化。方法设计特异性探针合成FAKSiRNA片段，脂质体法转染体外培养的增生性瘢痕成纤维细胞，光学显微镜观察细胞形态；MTT检测细胞活性并绘制细胞生长曲线；流式细胞仪检测细胞周期。结果转染后的成纤维细胞呈短梭形改变，胞质、突触减少，细胞核内核仁减少。细胞生长曲线表明．转染FAKsiRNA的瘢痕成纤维细胞生长明显抑制。转染后瘢痕成纤维细胞阻滞在G1期，S期和G2期细胞比例减少。结论FAK与瘢痕增生相关密切，因此改变FAK在成纤维细胞中的表达可望调控增生性瘢痕成纤维细胞的生物活性．

Effects of Specific Small Interfering RNA on Cell Proliferation of Human Hypertrophic Scar Fibroblasts

Objective To investigate the effect on a diverse array of cellular processes of human hypertrophic scar fibroblasts by using FAK specific small interfering RNA （SiRNA） on inhibiting FAK gene expression. Methods FAK siRNA was design with specific target finder and transfected into hypertrophic scar fibroblasts by the liposome-mediated gene transfection method. Light microscope was used to observe cell morphologic change. The viable cells were counted by MTF colorimetry and a cell growth curve was drawn. Flow cytometry was used to examine cell cycle. Results Abnormal change of cell morphology became into short fusiform, less cytoplasm synapses and less nucleoli in cell nucleolus after transfection. The cell growth curves indicated the prolifertation of cell trasfected with FAK specific SiRNA was inhibited significantly compared with controls. Transfected hypertrophic scar fibro blasts were blocked in G1 stage. The proportion of the cells decreased in S and G2 stage. Conclusion The quantity of FAK expression may has a significant inverse correlation with the forming of hypertrophic scar, The cellular process of hypertrophic scar fibroblasts can be controlled by regulating the expression of FAK.