​Earlier diagnosis and advances in treatment strategies have increased the average survival of cancer patients over the last decades. Despite the increased number of new anti-neoplastic agents, there has been no adequate therapy for intricate malignancies such as hepatic cancer. Cancer metabolism is the main building block standing behind cancer promotion and progression even in the presence of a harsh environment. Targeting metabolic pathways, such as glycolysis and metabolic regulators, is regarded as a promising new strategy for cancer treatment. The human hepatocellular carcinoma cell line, HepG2, was used to study the anti-proliferative activity of targeting PFKFB4 and HNF4-α mRNAs by transfection with siRNAs, each one alone and in combination. A conformational study was done to investigate the silencing of the above genes on their expression level using RT-PCR.The aim of the current study is to investigate the effect of knocking-down hepatic cancer glycolytic and metabolic regulators (PFKFB4, and HNF4-α) respectively, on cell’s viability using siRNA each one alone and in combination​​​