RhoA基因转染对QZG肝细胞生物学行为的影响及其机制的研究

目的探讨RhoA基因转染QZG肝细胞系后细胞骨架和体外侵袭能力的变化。方法构建持续激活型的RhoA真核表达载体,以脂质体介导转染肝细胞系QZG,用RT-PCR检测RhoA mRNA表达水平,观察细胞骨架的变化和体外侵袭能力。结果RT-PCR表明稳定转染细胞株中有RhoA mRNA高表达,而且在荧光显微镜下可见到细胞株绿色荧光,细胞形态和骨架发生变化。Boyden小室体外侵袭试验中实验组比空白对照组侵袭能力增加41.4%（P=0.006）。结论RhoA影响细胞的骨架变化和侵袭能力,在肝癌发生和发展中发挥重要作用。

Mechanism research and effects of transfection of RhoA gene on the biological behavior of human liver cell line QZG

Objective To investigate mechanism and effects of transfection of RhoA gene on the cytoskeleton change and biological behavior in QZG cell in vitro.Methods A human hepatocyte line stably expressing the dominant active mutation of RhoA gene,RhoA（G14V）,namely RhoAV14,was established.The mRNA expression levels of the constitutively activated form of RhoA were examined by RT-PCR analysis and were compared with those of the control cell line which was transfected with the plasmid pEGFP and the parent cell line QZG.The effects of RhoA protein on cytoskeleton and cell invasion were investigated to analyze the mechanism of RhoAV14 Expression on cell biological behaviors.Results RhoA expression was observed by RT-PCR and under fluorescence microscope.We found the morphological change of cells from spherical QZG cells to polygonal-shaped which RhoA activity was enhanced.Transwell invasion assay revealed that significantly higher RhoAV14 cells were able to migrate through the filters compared with mock cells（P=0.006）.Conclusions RhoA overexpression can significantly increase invasion of cells.RhoA may play an important role in invasion of tumor cells and likely to be a molecular target of blocking invasion.