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β-Elemene Inhibits Cell Proliferation by Regulating the Expression and Activity of Topoisomerases I and IIα in Human Hepatocarcinoma HepG-2 Cells.

Gong M, Liu Y, Zhang J, Gao YJ, Zhai PP, Su X, Li X, Li Y, Hou L, Cui XN - Biomed Res Int (2015)

Bottom Line: To investigate the effects of β-Elemene (β-ELE) on the proliferation, apoptosis, and topoisomerase I (TOPO I) and topoisomerase IIα (TOPO IIα) expression and activity of human hepatocarcinoma HepG-2 cells.After treatment with β-ELE, morphological alterations of HepG-2 cells were observed under an inverted microscope.Supercoiled pBR322 and kDNA were also used to determine the direct effect of β-ELE on DNA breaks. β-ELE significantly inhibited HepG-2 cell proliferation in a dose- and time-dependent manner. β-ELE also induced tumor cell arrest at S phase, induced cell apoptosis, and downregulated the protein expression of TOPO I and TOPO IIα in a dose-dependent manner. β-ELE also inhibited TOPO I- and TOPO IIα-mediated DNA relaxation but did not directly induce DNA breakage at any concentration. β-ELE could inhibit the proliferation of HepG-2 cells and interfere with the expression and activity of TOPO I and TOPO IIα.

View Article: PubMed Central - PubMed

Affiliation: Department of Oncology, The First Affiliated Hospital of Dalian Medical University, Dalian, Liaoning 116011, China.

ABSTRACT

Objective: To investigate the effects of β-Elemene (β-ELE) on the proliferation, apoptosis, and topoisomerase I (TOPO I) and topoisomerase IIα (TOPO IIα) expression and activity of human hepatocarcinoma HepG-2 cells.

Methods: After treatment with β-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 IIα was analyzed by Western blot techniques, and their activity was measured using the TOPO I-mediated, supercoiled pBR322 DNA relaxation and TOPO IIα-mediated Kinetoplast DNA (kDNA) decatenation assays, respectively. Supercoiled pBR322 and kDNA were also used to determine the direct effect of β-ELE on DNA breaks.

Results: β-ELE significantly inhibited HepG-2 cell proliferation in a dose- and time-dependent manner. β-ELE also induced tumor cell arrest at S phase, induced cell apoptosis, and downregulated the protein expression of TOPO I and TOPO IIα in a dose-dependent manner. β-ELE also inhibited TOPO I- and TOPO IIα-mediated DNA relaxation but did not directly induce DNA breakage at any concentration.

Conclusion: β-ELE could inhibit the proliferation of HepG-2 cells and interfere with the expression and activity of TOPO I and TOPO IIα.

No MeSH data available.


Related in: MedlinePlus

β-ELE induces cell cycle arrest and apoptosis in HepG-2 cells. (a) The effect of β-ELE (β-Elemene) on the cell cycle of HepG-2 cells after 24 h. (a1) Control group (without β-ELE). (a2) Experimental group (β-ELE 20 μg/mL). (a3) Experimental group (β-ELE 40 μg/mL). (a4) Experimental group (β-ELE 60 μg/mL). After treatment with different concentrations of β-ELE (0, 20, 40, and 60 μg/mL) for 24 h, a significant S phase arrest in HepG-2 cells was observed. The percentage of cells in S phase was 30.40 ± 1.19%, 39.38 ± 0.93%, and 51.43 ± 1.68% at 24 h when treated with β-ELE at 20, 40, and 60 μg/mL, respectively. These values were apparently higher than that of the control group after 24 h (18.29 ± 0.94%, P < 0.05). (b) The effect of β-ELE (β-Elemene) on the cell cycle of HepG-2 cells after 48 h. (b1) Control group (without β-ELE). (b2) Experimental group (β-ELE 20 μg/mL). (b3) Experimental group (β-ELE 40 μg/mL). (b4) Experimental group (β-ELE 60 μg/mL). After treatment with different concentrations of β-ELE (1, 20, 40, and 60 μg/mL) for 48 h, a significant S phase arrest in HepG-2 cells was observed. The percentage of cells in S phase was 42.36 ± 3.40%, 47.86 ± 4.83%, and 60.95 ± 4.61% after 48 h when treated with β-ELE at 20 μg/mL, 40 μg/mL, and 60 μg/mL, respectively. These values were apparently higher than that of the control group at 48 h (21.47 ± 0.59%, P < 0.05). (c) β-Elemene induces human hepatocarcinoma HepG-2 cell arrest at S phase. After treatment with different concentrations of β-ELE (0, 20, 40, and 60 μg/mL) for 24 h and 48 h, a significant S phase arrest in HepG-2 cells was observed. The percentage of cells in S phase was 30.40 ± 1.19%, 39.38 ± 0.93%, and 51.43 ± 1.68% after 24 h and 42.36 ± 3.40%, 47.86 ± 4.83%, and 60.95 ± 4.61% after 48 h when treated with β-ELE at 20, 40, and 60 μg/mL, respectively. These values were apparently higher than those of the control group at 24 h (18.29 ± 0.94%, P < 0.05) (Figure 4) and 48 h (21.47 ± 0.59%, P < 0.05) (Figure 5), which suggests that β-ELE treatment leads to an accumulation of HepG-2 cells in S phase (∗∗P < 0.005). (d) The effect of β-ELE (β-Elemene) on the apoptosis of human hepatocarcinoma HepG-2 cells. After treatment with different concentrations of β-ELE (20, 40, and 60 μg/mL) for 24 h and 48 h, the percentage of apoptosis cells was 11.94 ± 1.6%, 24.61 ± 2.07%, and 32.81 ± 2.58% after 24 h and 14.69 ± 1.77%, 27.14 ± 0.87%, and 34.38 ± 2.61% after 48 h. These values were significantly higher than those of the control group at 24 h (0.82 ± 0.27%, P < 0.05) and 48 h (1.07 ± 0.35%, P < 0.05) and showed dose and time dependence (∗∗P < 0.005).
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fig4: β-ELE induces cell cycle arrest and apoptosis in HepG-2 cells. (a) The effect of β-ELE (β-Elemene) on the cell cycle of HepG-2 cells after 24 h. (a1) Control group (without β-ELE). (a2) Experimental group (β-ELE 20 μg/mL). (a3) Experimental group (β-ELE 40 μg/mL). (a4) Experimental group (β-ELE 60 μg/mL). After treatment with different concentrations of β-ELE (0, 20, 40, and 60 μg/mL) for 24 h, a significant S phase arrest in HepG-2 cells was observed. The percentage of cells in S phase was 30.40 ± 1.19%, 39.38 ± 0.93%, and 51.43 ± 1.68% at 24 h when treated with β-ELE at 20, 40, and 60 μg/mL, respectively. These values were apparently higher than that of the control group after 24 h (18.29 ± 0.94%, P < 0.05). (b) The effect of β-ELE (β-Elemene) on the cell cycle of HepG-2 cells after 48 h. (b1) Control group (without β-ELE). (b2) Experimental group (β-ELE 20 μg/mL). (b3) Experimental group (β-ELE 40 μg/mL). (b4) Experimental group (β-ELE 60 μg/mL). After treatment with different concentrations of β-ELE (1, 20, 40, and 60 μg/mL) for 48 h, a significant S phase arrest in HepG-2 cells was observed. The percentage of cells in S phase was 42.36 ± 3.40%, 47.86 ± 4.83%, and 60.95 ± 4.61% after 48 h when treated with β-ELE at 20 μg/mL, 40 μg/mL, and 60 μg/mL, respectively. These values were apparently higher than that of the control group at 48 h (21.47 ± 0.59%, P < 0.05). (c) β-Elemene induces human hepatocarcinoma HepG-2 cell arrest at S phase. After treatment with different concentrations of β-ELE (0, 20, 40, and 60 μg/mL) for 24 h and 48 h, a significant S phase arrest in HepG-2 cells was observed. The percentage of cells in S phase was 30.40 ± 1.19%, 39.38 ± 0.93%, and 51.43 ± 1.68% after 24 h and 42.36 ± 3.40%, 47.86 ± 4.83%, and 60.95 ± 4.61% after 48 h when treated with β-ELE at 20, 40, and 60 μg/mL, respectively. These values were apparently higher than those of the control group at 24 h (18.29 ± 0.94%, P < 0.05) (Figure 4) and 48 h (21.47 ± 0.59%, P < 0.05) (Figure 5), which suggests that β-ELE treatment leads to an accumulation of HepG-2 cells in S phase (∗∗P < 0.005). (d) The effect of β-ELE (β-Elemene) on the apoptosis of human hepatocarcinoma HepG-2 cells. After treatment with different concentrations of β-ELE (20, 40, and 60 μg/mL) for 24 h and 48 h, the percentage of apoptosis cells was 11.94 ± 1.6%, 24.61 ± 2.07%, and 32.81 ± 2.58% after 24 h and 14.69 ± 1.77%, 27.14 ± 0.87%, and 34.38 ± 2.61% after 48 h. These values were significantly higher than those of the control group at 24 h (0.82 ± 0.27%, P < 0.05) and 48 h (1.07 ± 0.35%, P < 0.05) and showed dose and time dependence (∗∗P < 0.005).

Mentions: A significant S phase arrest in HepG-2 cells induced by β-ELE was observed (Figure 4). The percentage of cells in S phase was 30.40 ± 1.19%, 39.38 ± 0.93%, and 51.43 ± 1.68% after 24 h and 42.36 ± 3.40%, 47.86 ± 4.83%, and 60.95 ± 4.61% after 48 h, when treated with 20, 40, and 60 μg/mL of β-ELE, respectively. All of these values were apparently higher than those of the control group after 24 h (18.29 ± 0.94%, P < 0.05) (Figure 4(a)) and 48 h (21.47 ± 0.59%, P < 0.05) (Figure 4(b)), which suggests that β-ELE treatment leads to an accumulation of HepG-2 cells in S phase (Figure 4(c)).


β-Elemene Inhibits Cell Proliferation by Regulating the Expression and Activity of Topoisomerases I and IIα in Human Hepatocarcinoma HepG-2 Cells.

Gong M, Liu Y, Zhang J, Gao YJ, Zhai PP, Su X, Li X, Li Y, Hou L, Cui XN - Biomed Res Int (2015)

β-ELE induces cell cycle arrest and apoptosis in HepG-2 cells. (a) The effect of β-ELE (β-Elemene) on the cell cycle of HepG-2 cells after 24 h. (a1) Control group (without β-ELE). (a2) Experimental group (β-ELE 20 μg/mL). (a3) Experimental group (β-ELE 40 μg/mL). (a4) Experimental group (β-ELE 60 μg/mL). After treatment with different concentrations of β-ELE (0, 20, 40, and 60 μg/mL) for 24 h, a significant S phase arrest in HepG-2 cells was observed. The percentage of cells in S phase was 30.40 ± 1.19%, 39.38 ± 0.93%, and 51.43 ± 1.68% at 24 h when treated with β-ELE at 20, 40, and 60 μg/mL, respectively. These values were apparently higher than that of the control group after 24 h (18.29 ± 0.94%, P < 0.05). (b) The effect of β-ELE (β-Elemene) on the cell cycle of HepG-2 cells after 48 h. (b1) Control group (without β-ELE). (b2) Experimental group (β-ELE 20 μg/mL). (b3) Experimental group (β-ELE 40 μg/mL). (b4) Experimental group (β-ELE 60 μg/mL). After treatment with different concentrations of β-ELE (1, 20, 40, and 60 μg/mL) for 48 h, a significant S phase arrest in HepG-2 cells was observed. The percentage of cells in S phase was 42.36 ± 3.40%, 47.86 ± 4.83%, and 60.95 ± 4.61% after 48 h when treated with β-ELE at 20 μg/mL, 40 μg/mL, and 60 μg/mL, respectively. These values were apparently higher than that of the control group at 48 h (21.47 ± 0.59%, P < 0.05). (c) β-Elemene induces human hepatocarcinoma HepG-2 cell arrest at S phase. After treatment with different concentrations of β-ELE (0, 20, 40, and 60 μg/mL) for 24 h and 48 h, a significant S phase arrest in HepG-2 cells was observed. The percentage of cells in S phase was 30.40 ± 1.19%, 39.38 ± 0.93%, and 51.43 ± 1.68% after 24 h and 42.36 ± 3.40%, 47.86 ± 4.83%, and 60.95 ± 4.61% after 48 h when treated with β-ELE at 20, 40, and 60 μg/mL, respectively. These values were apparently higher than those of the control group at 24 h (18.29 ± 0.94%, P < 0.05) (Figure 4) and 48 h (21.47 ± 0.59%, P < 0.05) (Figure 5), which suggests that β-ELE treatment leads to an accumulation of HepG-2 cells in S phase (∗∗P < 0.005). (d) The effect of β-ELE (β-Elemene) on the apoptosis of human hepatocarcinoma HepG-2 cells. After treatment with different concentrations of β-ELE (20, 40, and 60 μg/mL) for 24 h and 48 h, the percentage of apoptosis cells was 11.94 ± 1.6%, 24.61 ± 2.07%, and 32.81 ± 2.58% after 24 h and 14.69 ± 1.77%, 27.14 ± 0.87%, and 34.38 ± 2.61% after 48 h. These values were significantly higher than those of the control group at 24 h (0.82 ± 0.27%, P < 0.05) and 48 h (1.07 ± 0.35%, P < 0.05) and showed dose and time dependence (∗∗P < 0.005).
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fig4: β-ELE induces cell cycle arrest and apoptosis in HepG-2 cells. (a) The effect of β-ELE (β-Elemene) on the cell cycle of HepG-2 cells after 24 h. (a1) Control group (without β-ELE). (a2) Experimental group (β-ELE 20 μg/mL). (a3) Experimental group (β-ELE 40 μg/mL). (a4) Experimental group (β-ELE 60 μg/mL). After treatment with different concentrations of β-ELE (0, 20, 40, and 60 μg/mL) for 24 h, a significant S phase arrest in HepG-2 cells was observed. The percentage of cells in S phase was 30.40 ± 1.19%, 39.38 ± 0.93%, and 51.43 ± 1.68% at 24 h when treated with β-ELE at 20, 40, and 60 μg/mL, respectively. These values were apparently higher than that of the control group after 24 h (18.29 ± 0.94%, P < 0.05). (b) The effect of β-ELE (β-Elemene) on the cell cycle of HepG-2 cells after 48 h. (b1) Control group (without β-ELE). (b2) Experimental group (β-ELE 20 μg/mL). (b3) Experimental group (β-ELE 40 μg/mL). (b4) Experimental group (β-ELE 60 μg/mL). After treatment with different concentrations of β-ELE (1, 20, 40, and 60 μg/mL) for 48 h, a significant S phase arrest in HepG-2 cells was observed. The percentage of cells in S phase was 42.36 ± 3.40%, 47.86 ± 4.83%, and 60.95 ± 4.61% after 48 h when treated with β-ELE at 20 μg/mL, 40 μg/mL, and 60 μg/mL, respectively. These values were apparently higher than that of the control group at 48 h (21.47 ± 0.59%, P < 0.05). (c) β-Elemene induces human hepatocarcinoma HepG-2 cell arrest at S phase. After treatment with different concentrations of β-ELE (0, 20, 40, and 60 μg/mL) for 24 h and 48 h, a significant S phase arrest in HepG-2 cells was observed. The percentage of cells in S phase was 30.40 ± 1.19%, 39.38 ± 0.93%, and 51.43 ± 1.68% after 24 h and 42.36 ± 3.40%, 47.86 ± 4.83%, and 60.95 ± 4.61% after 48 h when treated with β-ELE at 20, 40, and 60 μg/mL, respectively. These values were apparently higher than those of the control group at 24 h (18.29 ± 0.94%, P < 0.05) (Figure 4) and 48 h (21.47 ± 0.59%, P < 0.05) (Figure 5), which suggests that β-ELE treatment leads to an accumulation of HepG-2 cells in S phase (∗∗P < 0.005). (d) The effect of β-ELE (β-Elemene) on the apoptosis of human hepatocarcinoma HepG-2 cells. After treatment with different concentrations of β-ELE (20, 40, and 60 μg/mL) for 24 h and 48 h, the percentage of apoptosis cells was 11.94 ± 1.6%, 24.61 ± 2.07%, and 32.81 ± 2.58% after 24 h and 14.69 ± 1.77%, 27.14 ± 0.87%, and 34.38 ± 2.61% after 48 h. These values were significantly higher than those of the control group at 24 h (0.82 ± 0.27%, P < 0.05) and 48 h (1.07 ± 0.35%, P < 0.05) and showed dose and time dependence (∗∗P < 0.005).
Mentions: A significant S phase arrest in HepG-2 cells induced by β-ELE was observed (Figure 4). The percentage of cells in S phase was 30.40 ± 1.19%, 39.38 ± 0.93%, and 51.43 ± 1.68% after 24 h and 42.36 ± 3.40%, 47.86 ± 4.83%, and 60.95 ± 4.61% after 48 h, when treated with 20, 40, and 60 μg/mL of β-ELE, respectively. All of these values were apparently higher than those of the control group after 24 h (18.29 ± 0.94%, P < 0.05) (Figure 4(a)) and 48 h (21.47 ± 0.59%, P < 0.05) (Figure 4(b)), which suggests that β-ELE treatment leads to an accumulation of HepG-2 cells in S phase (Figure 4(c)).

Bottom Line: To investigate the effects of β-Elemene (β-ELE) on the proliferation, apoptosis, and topoisomerase I (TOPO I) and topoisomerase IIα (TOPO IIα) expression and activity of human hepatocarcinoma HepG-2 cells.After treatment with β-ELE, morphological alterations of HepG-2 cells were observed under an inverted microscope.Supercoiled pBR322 and kDNA were also used to determine the direct effect of β-ELE on DNA breaks. β-ELE significantly inhibited HepG-2 cell proliferation in a dose- and time-dependent manner. β-ELE also induced tumor cell arrest at S phase, induced cell apoptosis, and downregulated the protein expression of TOPO I and TOPO IIα in a dose-dependent manner. β-ELE also inhibited TOPO I- and TOPO IIα-mediated DNA relaxation but did not directly induce DNA breakage at any concentration. β-ELE could inhibit the proliferation of HepG-2 cells and interfere with the expression and activity of TOPO I and TOPO IIα.

View Article: PubMed Central - PubMed

Affiliation: Department of Oncology, The First Affiliated Hospital of Dalian Medical University, Dalian, Liaoning 116011, China.

ABSTRACT

Objective: To investigate the effects of β-Elemene (β-ELE) on the proliferation, apoptosis, and topoisomerase I (TOPO I) and topoisomerase IIα (TOPO IIα) expression and activity of human hepatocarcinoma HepG-2 cells.

Methods: After treatment with β-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 IIα was analyzed by Western blot techniques, and their activity was measured using the TOPO I-mediated, supercoiled pBR322 DNA relaxation and TOPO IIα-mediated Kinetoplast DNA (kDNA) decatenation assays, respectively. Supercoiled pBR322 and kDNA were also used to determine the direct effect of β-ELE on DNA breaks.

Results: β-ELE significantly inhibited HepG-2 cell proliferation in a dose- and time-dependent manner. β-ELE also induced tumor cell arrest at S phase, induced cell apoptosis, and downregulated the protein expression of TOPO I and TOPO IIα in a dose-dependent manner. β-ELE also inhibited TOPO I- and TOPO IIα-mediated DNA relaxation but did not directly induce DNA breakage at any concentration.

Conclusion: β-ELE could inhibit the proliferation of HepG-2 cells and interfere with the expression and activity of TOPO I and TOPO IIα.

No MeSH data available.


Related in: MedlinePlus