Hepatocellular carcinoma (HCC) is a complicated disease with a poor prognosis and high mortality rates. The prevention, control, diagnosis, and treatment of liver cancer have become vital focuses in healthcare research. This study aimed to evaluate the in vitro effect of taurine (Tau) on the expression of miR-122-5p that targets some limiting glycolytic enzymes and affects the overall glycolytic pathway in HepG2 cells. IC50 and the inhibitory effect of Tau on cell proliferation were measured after 48h by MTT assay. Then, themRNA expressions of some apoptosis-relatedgenes P53, BAX, Caspase-3, and Bcl-2 weremeasured using quantitativereal-time (qRT-PCR) and theprotein levels were confirmed by enzyme-linked immunosorbentassay(ELISA). The activities of some antioxidant'sbiomarkers were assessed. The gene expression of miR-122-5p that targetssome limiting glycolytic enzymes; Aldolase and Lactate dehydrogenase (LDH), wereevaluated after treatment with Tau for 48h. A Significant inhibition in the proliferation of HepG2 was encountered after treatment with Tau in a dose-dependent manner. Moreover, the expression of apoptotic genes p53, Bax, and Caspase-3 exhibited a significant upregulation, while Bcl-2 showed a significant downregulation. These alterations in the expression levels were also confirmed on the protein level. The antioxidant activities of GPx, CAT, and NO were significantly elevated versus untreated control. Also, a significant increase in the expression level of miR-122-5p was observed after treatment with Tau affecting the metabolic activity of HCC cells. Concomitantly, a significant inhibition in ALDOA protein and the hallmark ofglycolytic enzymes LDH and Aldolase were observed. These observations showed that taurine inhibits HepG2 cell proliferation and restores the expression of miR-122-5p which inhibits the hallmark glycolytic enzymes and ultimately the metabolic activity of HCC cells. Tau is assumed to be a promising and effective antitumor therapy of HCC.