Materials and Methods…
Results and Discussion
The experimental results from the cell viability assay are shown below. The relative cell viabilities are summarized separately for each group of cells treated with different transfection reagents.
Cells treated with the transfection reagents alone typically yielded relative viabilities that were similar to the untreated cells in the case of transfection reagents Prime-Fect and ALL-Fect, indicating no effect of these transfection reagents alone on the cell viabilities. InVivo RNA-Fect and RJH-X was not tested in this regard. Cells treated with scrambled siRNA complexes of Prime-Fect and InVivo RNA-Fect gave relative viabilities equivalent to untreated cells, again indicating no toxicity of the non-specific complexes on the viabilities of A549 cells. Complexes from ALL-Fect and RJH-X displayed some loss of viabilities (~10%) indicating slight toxicity of the non-specific complexes compared to un-treated cells. In contrast, the cytotoxic siRNA complexes gave significant reduction of toxicity, with Prime-Fect complexes displaying ~40% relative cell viability and InVivo RNA-Fect complexes displaying ~16% relative cell viability, and ALL-Fect and RJH-X complexes displaying less than 10% relative cell viabilities. Therefore, multiple RJH reagents were found to be very effective in this particular application intended to induce death of lung cancer cells. The differential activity obtained between the scrambled siRNA and cytotoxic siRNA were very high, suggesting that effective therapeutic effects could be readily demonstrated. A relatively low concentration of siRNAs was used (50 nM) to obtain the desired therapeutic effect so that translation into animal models should be facilitated at such low doses.
Figure 1. Relative cell viability