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目的 探讨肝细胞生长因子(hepatocyte growth factor, HGF)基因对肝细胞生长增殖能力的影响。方法 通过脂质体介导法,将 HGF基因导入肝细胞中。用荧光显微镜以及原位杂交观察到HGF基因表达。采用检测细胞生长曲线、Ki 67蛋白和嗜银蛋白免疫组化观察肝细胞生长增殖能力及DNA合成能力的变化。结果 荧光显微镜观察可见到绿色荧光蛋白的表达,用原位杂交方法进一步证实了HGF蛋白在细胞中的表达。细胞生长曲线显示,转染HGF基因的肝细胞增殖速度明显增快,Ki 67蛋白和嗜银蛋白表达明显增多,提示转导 HGF基因使肝细胞增殖活性增加。结论 本实验显示转染的HGF可表达并有促细胞分裂活性。为进一步了解 HGF分子生物学特性及治疗应用提供了理论基础。
Objective To investigate the effect of hepatocyte growth factor (HGF) gene on the growth and proliferation of hepatocytes. Methods HGF gene was introduced into hepatocytes by liposome-mediated method. HGF gene expression was observed by fluorescence microscopy and in situ hybridization. Cell growth curve, Ki 67 protein and argyrophilic protein immunohistochemistry were used to observe the changes of hepatocyte proliferation and DNA synthesis. Results The expression of green fluorescent protein (GFP) was observed by fluorescence microscopy. The expression of HGF protein was further confirmed by in situ hybridization. The cell growth curve showed that the proliferation of hepatocytes transfected with HGF was significantly increased, and the expressions of Ki 67 and argyrophilic proteins were significantly increased, suggesting that transduction of HGF gene increased the proliferation of hepatocytes. Conclusion This experiment shows that transfected HGF can express and have mitogenic activity. Provide a theoretical basis for further understanding of the molecular biological characteristics of HGF and its therapeutic application.