醋酸棉酚对人舌鳞癌Cal-27细胞侵袭性作用的实验研究

Effect of Gossypol Acetate Acid on Invasion of Human Tongue Squamous Cell Carcinoma Cal-27 Cells in vitro

目的研究醋酸棉酚(gossypol acetic acid,GAA)对体外培养的人舌鳞癌Cal-27细胞侵袭影响及其作用机制。方法制备0μmol/L、2.5μmol/L、5.0μmol/L、10.0μmol/L的GAA作用于人舌鳞癌Cal-27细胞,通过Transwell实验检测GAA对人舌鳞癌Cal-27细胞侵袭能力的影响;通过qRT-PCR及Western blot实验检测GAA对人舌鳞癌Cal-27细胞基质金属蛋白酶-2(Matrix Metalloproteinase-2,MMP-2)、基质金属蛋白酶-9(Matrix Metalloproteinase-9,MMP-9)mRNA及蛋白的表达变化。结果Transwell实验显示:GAA对人舌鳞癌Cal-27细胞细胞侵袭产生抑制作用,实验组和对照组比较差异有统计学意义(P<0.05);qRT-PCR实验及Western blot显示,实验组人舌鳞癌Cal-27细胞中MMP-2、MMP-9 mRNA及蛋白的表达降低,与对照组相比差异有统计学意义(P<0.05)。结论GAA能抑制人舌鳞癌Cal-27细胞的侵袭性,并降低人舌鳞癌Cal-27细胞MMP-2、MMP-9的mRNA及蛋白表达水平,这可能是GAA抑制肿瘤侵袭作用的机制之一。

Objective To observe the effect of gossypol acetate acid(GAA)on the invasion of human tongue squamous cell carcinoma Cal-27 cells in vitro.Methods GAA of different concentrations(0μmol/L,2.5μmol/L,5.0μmol/L,10.0μmol/L)was acted on Cal-27 cells,Transwell experiment was employed to detect the inhibitory effect of GAA on the invasion of Cal-27 cells.The effect of gossypol acetate on MMP-2 and MMP-9 mRNA expression was quantified by qRT-PCR.The effect of gossypol acetate acid on MMP-2 and MMP-9 protein expression was analyzed by Western-Blot experiment.Results Cell Transwell chamber experiment showed that GAA could inhibit the migration of human tongue squamous cell carcinoma Cal-27 cells,and the amount of invasion decreased with the increase in concentration.The difference between the control group and the experimental group was statistically significant(P<0.05).qRT-PCR experiment and Western-blot results showed that the expression of MMP-2 and MMP-9 mRNA and proteins was significantly reduced after Cal-27 cells of human tongue squamous cell carcinoma was treated with GAA,and the difference between the control group and the experimental group was statistically significant(P<0.05).Conclusions GAA can reduce the invasiveness of tongue squamous cell carcinoma Cal-27 cells.And GAA can reduce the mRNA and protein expression levels of MMP-2 and MMP-9 of human tongue squamous cell carcinoma Cal-27 cells.