ABSTRACT
In order to study the important factors involved in cationic liposome-mediated gene transfer, Lipofectamine 2000 or DOTAP was evaluated using three types of cells (Hep-2, MCF-7 and SW-480) in vitro transfection efficiencies. Different properties of the two reagents were analyzed and compared by DNA arrearage assay and MTT assay. Both Lipofectamine 2000 and DOTAP had strong capability to combine with DNA; Lipofectamine 2000 can get higher transfection efficiency of the three cells by using GFP as report gene, meanwhile, DOTAP can also get higher transfection efficiency against Hep-2 cell. However, DOTAP showed lower transfection efficiency against MCF-7 and SW-480 cell. On the other hand, the cytotoxicity assay showed that over 85% cell viability of MCF-7 cell could be achieved both by Lipofectamine 2000 and DOTAP under the optimal transfection condition. Relatively speaking, Lipofectamine 2000 has very high transfection efficiency in a broad range of cell lines, but because of the special selectivity of cell type on liposome, DOTAP also has a broad application prospect.
Subject(s)
Humans , Cell Line, Tumor , Cell Survival , DNA , Genetics , Fatty Acids, Monounsaturated , Chemistry , Toxicity , Gene Transfer Techniques , Genes, Reporter , Genetic Vectors , Green Fluorescent Proteins , Metabolism , Lipids , Chemistry , Toxicity , Quaternary Ammonium Compounds , Chemistry , Toxicity , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To investigate the apoptosis and proliferation activity of Tca8113 cells.</p><p><b>METHODS</b>The vector that involves short hairpin RNA of iNOS was transfected to Tca8113 cells. The change of iNOS expression was observed using immunohistochemistry technique, the apoptosis rate examined by flow cytometry, and the proliferation Tca8113 cells examined by methyl thiazolyl tetrazolium (MTT).</p><p><b>RESULTS</b>The expression of iNOS in Psilencer-iNOS group was lower than that in control groups (P < 0.01), the apoptosis rate was higher than that in control groups (P < 0.01); whereas the proliferation activity of Tca8113 cells in Psilencer-iNOS group was lower than that in control groups.</p><p><b>CONCLUSIONS</b>Down expression of iNOS by RNAi can promotes apoptosis of Tca8113 cells and has an anti-proliferation activity effect.</p>