Objective. To investigate the effects of [beta]-Elemene ([beta]-ELE) on the proliferation, apoptosis, and topoisomerase I (TOPO I) and topoisomerase II[alpha] (TOPO II[alpha]) expression and activity of human hepatocarcinoma HepG-2 cells. Methods. After treatment with [beta]-ELE, morphological alterations of HepG-2 cells were observed under an inverted microscope. Cell proliferation was assessed using an MTT assay, cell cycles were analyzed using flow cytometry, and apoptosis was detected by Annexin V/PI staining. The expression of TOPO I and TOPO II[alpha] was analyzed by Western blot techniques, and their activity was measured using the TOPO I-mediated, supercoiled pBR322 DNA relaxation and TOPO II[alpha]-mediated Kinetoplast DNA (kDNA) decatenation assays, respectively. Supercoiled pBR322 and kDNA were also used to determine the direct effect of [beta]-ELE on DNA breaks. Results. [beta]-ELE significantly inhibited HepG-2 cell proliferation in a dose- and time-dependent manner. [beta]-ELE also induced tumor cell arrest at S phase, induced cell apoptosis, and downregulated the protein expression of TOPO I and TOPO IIa in a dose-dependent manner. [beta]-ELE also inhibited TOPO I- and TOPO II[alpha]-mediated DNA relaxation but did not directly induce DNA breakage at any concentration. Conclusion. [beta]-ELE could inhibit the proliferation of HepG-2 cells and interfere with the expression and activity of TOPO I and TOPO II[alpha].