Hexokinase (HK) is particularly identified as a key regulator in cancer metabolism and progression, especially the isoform hexokinase II (HK2). It facilitates a phenomenon known as the Warburg effect. This is the high glucose uptake and glycolytic flux typical of cancer cells, done by catalyzing the first committed step of glycolysis, that is, converting glucose to glucose-6-phosphate. HK2 is frequently overexpressed in a variety of cancers and is associated with the outer mitochondrial membrane, which promotes cell survival by inhibiting apoptotic signaling. Due to the dual role in the production of energy and apoptosis avoidance positions, HK2 is an interesting target for therapeutic intervention. The inhibition of the enzyme hexokinase by small-molecule inhibitors, RNA interference, clustered regularly interspaced short palindromic repeats (CRISPR/Cas9) gene editing, and metabolic pathway blockers has been observed in preclinical studies for their potential to decrease tumor growth, sensitize cancerous cells to radiation, as well as for chemotherapy and restricting metastasis. Despite encouraging outcomes, challenges like isoform selectivity, metabolic adaptation, and drug delivery remain. Continuous exploration of HK’s role in cancer biology and the development of tailored inhibitory techniques might result in more effective and less harmful cancer therapies.