The Initial Study On The Effects Of Genistein On The Proliferation And Invasion Of Human Tongue Carcinoma Cell Line Tca8113 In Vitro

Objective: To study the effects of Genistein on the proliferation and invasion of human tongue carcinoma cell line Tca8113 in vitro,and to approach tis mechanism of action.Methods: Human tongue carcinoma cell line Tca8113 was used in vitro as a study object. Different concentrations of Genistein were used as the exerting factors, and 0.1% DMSO as the control group. The effects of Genistein on the proliferation of Tca8113 were measured with MTT assay. The rate of apoptosis was detected with Annexin V/PI staining and the cell cycle was assayed by flow cytometry. The effects on Bcl-2,Bax and caspase-3 levels of Tca8113 were detected by flow cytometry. Effects of migration and invasion were assayed with the methods of transwell migration assay and scrape wound healing assay. The level of MMP-2 and activated MMP-2 in the ecto-medium were detected by genlatin zymography and Gel Imaging System. The MMP-2 level of Tca8113 cells was detected by immuocytochemistry and western blotting.Results:1: Genistein inhibited the proliferation of human tongue carcinoma cell line (Tca8113) significantly (P<0.05). The most potent inhibitory effect on proliferation occurred in 15μg/mL(P<0.01)genistein group. Genistein inhibited Tca8113 cell proliferation, and the inhibition ratio was in a concentration-dependent fashion with time and concentration.2:Flow cytometry displayed that Genistein(15μg/mL)induced the cell apoptosis at both 24h and 48h(P<0.05). The cell population of S phase did not change at 24h but was decreased significantly at 48h. The cell cycle was arrested in G2 transition,and ratio of G2/G1 was waxed.3:Flow cytometry displayed that Genistein decreased the expression of Bcl-2 in Tca8113,and increased the expression of Bax,caspase-3. The expression level was in a concentration-dependent fashion.4:Transwell migration assay and scrape wound healing assay displayed that more inhibitory effects on invasion occurred in 15μg/mL Genistein group compared with the control group(P<0.01).5:Genistein decreased the expression of MMP-2 by 28.31% and activated MMP-2 by 18.75% in the ecto-medium. Gensitein significantly inhibited the expression of MMP-2 in Tca8113 cells by the method of immunocytochemistry and western blotting.Conclusions:1:Genistein significantly inhibits the proliferation of Tca8113 and promotes its apoptosis. Genistein can change the cycle of cell, decreasing the the cell population of the S phase and arresting the cell cycle in G2 transition.2:The effect of Genistein to promote the apoptosis of Tca8113 has relationship with the decreased level of bcl-2 and the increased level of bax. The change of expression of bcl-2 and bax may have relationship with the signal channel of Genistein inducing apoptosis. 3:Genistein significantly depresses the migration and invasion of human tongue carinoma cell line Tca8113 in vitro and decreases the MMP-2 expression intracellularly and extracellularly.