Piezo1 is identified as a novel mechanosensitive ion channel protein regulating a variety of physiological and pathological processes. However, the cellular effects of activation of Piezo1 protein on hepatocellular carcinoma (HCC) remains to be illuminated. In this study, we investigated the interaction between Piezo1 activation associated with metabolic reprogramming, pro-inflammatory cytokine release, and HCC progression. Our results indicated that the expression of Piezo1 gene and protein was up-regulated in the cancerous tissues of HCC patients, and the increased mRNA levels of Piezo1 were associated with unfavorable clinical outcomes. The activation of Piezo1 by its agonist Yoda1, facilitated HCC development by stimulating proliferation, migration, and invasion. Furthermore, Piezo1 channel activation by its agonist mediated pro-inflammatory responses mainly through its downstream molecule CXCL8 via ERK1/2 and AKT signaling pathways by up-regulating CXCL8 expression in transcriptional levels of HCC cells. The secretion of CXCL8 was induced by Piezo1 activation in HCC cells, improving immune cells trafficking and promoting angiogenesis, and therefore cross-talked with the tumor microenvironment (TME). Additionally, increased expression of MTHFD2, a metabolic checkpoint enzyme, was induced by the activation of Piezo1 channel and was likely to be a downstream target involved in mitochondrial stress followed by Piezo1 channel activation. In vitro data was further corroborated by the deceleration of tumor growth sizes in nude mice subcutaneously injected with Piezo1-depleted HCC cells in vivo. Finally, we showed synergistic anti-tumor effects of GsMTx4 and arachidonic acid, two Piezo1 antagonists, together with oxaliplatin in HCC cells. In summary, our findings suggest that the present linkage of metabolic reprogramming and pro-inflammatory responses after Piezo1 activation in HCC. Targeting Piezo1 appears to be a novel therapeutic strategy improving the treatment efficacy in HCC.