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.