Hepatocellular carcinoma (HCC) is a liver malignancy which lacks effective treatment and with poor prognosis. β-Elemene refers to a series of natural Curcuma wenyujin-derived compounds, exerting lots of biological activities, which is especially famous for it's antitumor properties.Exploring the underlying mechanism of β-Elemene against HCC.MTT, the assay of Colony formation and Flow cytometric were employed to evaluate the growth of HCC and LO2 cells by β-Elemene. HOTAIR、SP1 and PDK1 plasmids were transfected into HCC cells by transient transfection assay, and the expression and interaction of HOTAIR、SP1 and PDK1 were assessed via qRT-PCR and Western Blotting.β-Elemene suppressed HCC cell growth through downregulating HOTAIR, SP1 and PDK1. Mechanism experiments proved that there existed reciprocal interaction among HOTAIR, SP1 and PDK1. Exogenously overexpressed HOTAIR or SP1 eliminated the suppressive properties of β-Elemene on them, and both of which regulated PDK1 expression in HCC cells. Additionally, exogenously overexpressed SP1 or HOTAIR prevented β-Elemene inhibition of the protein-level expression of PDK1, whereas overexpressing PDK1 had no effect on SP1, though it still weakened the inhibition of cell growth and HOTAIR expression by β-Elemene.β-Elemene suppresses HCC cell proliferation via through the regulation of HOTAIR/SP1/PDK1 axis and their interaction.Copyright © 2021. Published by Elsevier B.V.