Evaluation of efficacy of apoptosis-inducing gene therapy in HCT116 colorectal cancer cells
Soňa Bálintová 1
Silvia Tyčiaková 2
Miroslava Matúšková 2
1 Department of Genetics, Faculty of Natural Sciences, Comenius University, Bratislava 2 Cancer Research Institute, Biomedical Research Center, Slovak Academy of Sciences, Bratislava
|Section:||Cellular metabolism, physiology, molecular biology and genetics|
Colorectal cancer remains one of the leading causes of cancer-related death worldwide. The treatment outcome for the advanced stages of the disease is unfavorable, thus the design of new therapeutic modalities is of high importance. The programming of cancer cells to undergo apoptosis, triggered by the introduction of a suicide gene, can represent a promising approach. A major candidate, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), induces both the extrinsic as well as the intrinsic pathway of apoptosis.
We decided to use a gene therapy approach to introduce a recombinant TRAIL gene into the cell, wherefrom it will be transcribed and translated as a functional ligand. Using molecular biology techniques, we prepared an expression vector coding for the human TRAIL gene. We identified three positive bacterial clones carrying the newly prepared pCIneo-TRAIL plasmid. pCIneo-GFP plasmid served as a control vector. The transfection of the HCT116 cell line with the pCIneoGFP vector led to an induction of green fluorescence 24 hours post-transfection , which proves the efficacy of the control vector. 72 hours after the transfection of the HCT116 cell line with the pCIneo-TRAIL plasmid, we observed a statistically significant increase in the proportion of apoptotic as well as overall dead cells in the culture compared to non-transfected cells.
Our results suggest that TRAIL acts as an apoptosis-inducing agent and may serve as a candidate for the treatment of colorectal cancer cells. Further experiments are required to optimize the timing design of transfection and apoptosis detection with the aim to achieve a lowered late apoptotic/necrotic and enhanced early apoptotic rate.