Effect Of Gossypol Acetate Acid On Migration And Invasion Of Cal-27 Cells In Human Tongue Squamous Cell Carcinoma

Objective:1.To observe the effect of gossypol acetate acid(GAA)on the invasion and migration of human tongue squamous cell carcinoma Cal-27 cells in vitro;to explore the mechanism of gossypol acetic acid's effect on the invasion of human tongue squamous cell carcinoma Cal-27 cells in hope to lay relevant experimental and theoretical foundations for clinical tumor treatment.Methods:1.GAA of different concentrations(2.5?mol/L?5.0?mol/L?10.0?mol/L)was dissolved in dimethyl sulfoxide(DMSO)to act on Cal-27 cells,and complete medium with DMSO was prepared and medium without GAA as the control group.The experimental time points were set as 3h,6h,9h,12h,15h,18h,21h and 24h.2.The effect of GAA on the migration ability of Cal-27 cells was studied with scratch test.3.Transwell chamber experiment was employed to detect the inhibitory effect of GAA on the invasion and the migration of Cal-27 cells.4.The effect of gossypol acetate on MMP-2 and MMP-9 mRNA expression was quantified by qRT-PCR.5.The effect of gossypol acetate acid on MMP-2 and MMP-9 protein expression was analyzed by Western-blot experiment.Results:1.cell scratch test showed that GAA with concentrations of 2.5?mol/L,5.0?mol/L and 10.0?mol/L could inhibit the migration of human tongue squamous cell carcinoma Cal-27 cells,and within the experimental concentration range,the amount of migration decreased with the increase in concentration.The difference between the control group and the experimental group was statistically significant(P<0.05),suggesting that GAA could effectively inhibit the migration of Cal-27 cells.2.The results of transwall experiment show that GAA with concentrations of 2.5?mol/L,5.0?mol/L and 10.0?mol/L has certain inhibitory effects on the invasion and migration of Cal-27 cells,and within the experimental concentration range,its invasion ability decreased with the increase in GAA concentration.The difference between the control group and the experimental group was statistically significant(P<0.05).3.qRT-PCR experiment showed that the expression of MMP-2 and MMP-9 mRNA was significantly reduced after Cal-27 cells of human tongue squamous cell carcinoma was treated with GAA of 2.5?mol/L,5.0?mol/L,and 10.0?mol/L.Moreover,the higher the GAA concentration,the lower the mRNA expression of MMP-2 and MMP-9.And the difference between the control group and the experimental group was statistically significant(P<0.05).4.Western-blot results showed that the relative expression of matrix metalloproteinase-2 and matrix metalloproteinase-9(MMP-2,MMP-9)proteins decreased 24 hours after GAA with 2.5?mol/L,5.0?mol/L,and 10.0?mol/L concentrations acted on human tongue squamous cell carcinoma Cal-27 cells,and the decrease increased significantly with the increase in GAA concentration.The difference between the control group and the experimental group was statistically significant(P<0.05).Conclusions:1.GAA can reduce the mobility and invasiveness of Cal-27 cells of tongue squamous cell carcinoma.In the experimental concentration range,the mobility and invasiveness decreased with the increase in GAA concentration and acting time.2.A certain concentration of GAA can reduce the mRNA and protein expression levels of MMP-2 and MMP-9 in Cal-27 cells of human tongue squamous cell carcinoma.