Next Generation Sequencing Facilitates Quantitative Analysis of HCC cells transfected with NCsiRNA or ODC1siRNA

We investigated if targeting of ODC1 with siRNA could inhibit HCC progression. To accomplish this, Huh1 and Huh7 human HCC cells were transfected with ODC1 siRNA and tested for growth inhibition and apoptotic induction using MTS, FACS and microscopic analysis. To obtain insights into the molecular changes in response to ODC1 knockdown, global changes in gene expression were examined using RNA sequencing. Our results show that ODC1 silencing caused inhibition of HCC cell growth through blockade of cell cycle progression and a strong induction of apoptosis. Subsequent Western blotting revealed that ODC1 silencing decreased the levels of cell cycle regulator cyclin E1, PARP-1, and pro-caspase 3 accelerating apoptotic signaling in HCC cells. In addition, ODC1 silencing resulted in a strong inhibition in the expressions of important regulators of glycolysis and lipogenesis, GLUT4 and ChREBP, and caused a marked decrease in fat accumulation. These findings suggest that ODC1 can be a promising target for systemic therapy of HCC by modulating tumor metabolism. Overall design: Examination of 2 different siRNA transfected cells in 2 cell lines.