醋酸棉酚对人舌鳞癌Tca8113细胞hMLH1蛋白及mRNA表达的影响

Effects of Gossypol Acetic Acid on Protein and mRNA Expressions of hMLH1 Gene in Human Tongue Carcinoma Cell Line Tca8113

目的研究醋酸棉酚(gossypol acetic acid,GAA)对人舌鳞癌Tca8113细胞hMLH1蛋白及mRNA表达的影响,探讨GAA的抗肿瘤作用机制.方法 (1)应用Western-blot法检测Tca8113细胞在GAA作用48 h后hMLH1基因的蛋白表达变化;(2)应用实时荧光定量(RFQ-PCR)法检测Tca8113细胞在GAA作用48 h后hMLH1基因mRNA的表达变化.结果 (1)Western-blot检测显示:分别用终浓度5μmol/L、10μmol/L、15μmol/L的GAA作用人舌鳞癌Tca8113细胞48 h后,hMLH1基因的蛋白表达比对照组明显增高(P<0.05);(2)RFQ-PCR检测显示:以终浓度5μmol/L、10μmol/L、15μmol/L的GAA作用于Tca8113细胞48 h后hMLH1基因的mRNA表达水平高于对照组(P<0.05).结论一定浓度的GAA能够使人舌鳞癌Tca8113细胞系hMLH1基因的蛋白及mRNA表达上调,这可能是GAA抗肿瘤作用机制之一.

Objective The purpose of this study was to investigate the effects of gossypol acetic acid （GAA） on protein and mRNA expressions of hMLH1 gene in human tongue carcinoma cell line Tca8113 in vitro in order to discuss the mechanism of tumor suppression of GAA. Methods （1） Western-blot was used to study the effects of GAA on protein expressions of hMLH1 gene in Tca8113 cell line treated by different concentrations of GAA for 48 h. （2） Real-time fluorescence quantitative PCR （RFQ-PCR） was used to investigate the effects on the mRNA expressions of hMLH1 gene in Tca8113 cell line treated by GAA for 48 h. Results （1） Compared with the control group, the results of Western-blot showed that the protein expression of hMLH1 gene was increased after treatment by GAA for 48 h （P 〈0.05） . （2） The results of RFQ-PCR indicated that the mRNA expression of hMLH1 gene was increased after GAA treatment for 48 h （ 〈0.05） . Conclusion GAA could up-regulate protein and mRNA expression of hMLH1 in Tca8113 cell line, which indicated that it may be one of the mechanisms of tumor suppression effect of GAA.