Effect of Genistein on Apoptosis and Proliferation of Hepatocellular Carcinoma Hepa1-6 Cell Line

Abstract

Background: One of the main causes of mortality is hepatocellular carcinoma (HCC) which accounts for the third leading cause of deaths and one in forty deaths worldwide. The flavonoids, natural antioxidant compounds, account for a major group of polyphenolic compounds. One of the major isoflavones in soybean is genistein (GE) which can inhibit proliferation and induce apoptosis. Isoflavones, major type of phenolic materials, derived from dietary plants and medicinal herbs play a significant role in cancer prevention and treatment. Correlation between dietary habits and cancer risk including breast, prostate, and colon cancer has been reported. Various bioactivities of these compounds such as anticarcinogenic and antioxidant are responsible for their chemopreventive activities by which induce migration, proliferation, cell cycle arrest, and apoptosis. GE, one of the major isoflavones, is considered as a potent chemopreventive agent against cancer. The aim of this study was to investigate the inhibitory and apoptotic effects of GE on HCC Hepa1-6 cell line.

Methods: Cell viability assay and cell cycle analysis with flow cytometry were used to evaluate proliferative and apoptotic effect GE.

Results: GE inhibited the growth of Hepa1-6 cells and induced apoptosis with a concentration and time-dependent fashion. During GE treatment for 24, the half maximal inhibitory concentration (IC50) was 20 μM, and the maximum inhibition of cell growth was 52% (P < 0.01). The percentage of apoptotic cells with a concentration of 20 μM of GE after 24, 48, and 72 h was 35, 42, and 65%, respectively (P < 0.01).

Conclusions: Our finding clearly indicated that GE can significantly inhibit proliferation of hepatocellular carcinoma Hepa 1-6 cell line and induce apoptosis in this cell line.

Keywords: Apoptosis; genistein; hepatocellular carcinoma; proliferation.

Figure 1

Inhibitory effect of genistein on proliferation of Hepa1-6. Hepa1-6 cells were treated with various concentration of genistein (as indicated in graph). The data are presented as the percentage change compared with the control group. *P < 0.001, significantly different from the control group

Figure 2

Antiproliferative effects of genistein (20 μM) on Hep1-6 cells in different time periods (24, 48, and 72 h). The data are presented as the percentage change compared with the control group. *P < 0.001, significantly different from the control group

Figure 3

Effects of genistein on Hepa1-6 cell apoptosis. The cells were treated with genistein (20 μM) for 24, 48, and 72 h and the apoptosis inducing effect of genistein was investigated by flow cytometric analysis of Hepa1-6 cells stained with Annexin V and propidium iodide. Results were obtained from three independent experiments and were expressed as mean ± standard error of the mean. P < 0.001