目的:研究VES对Her-2、p53共表达乳腺癌细胞株MDA-MB-453细胞的增殖、凋亡的影响、通过检测Her-2、p53探讨VES抑制乳腺癌细胞生长的机制.方法:应用 MTT法检测VES对MDA-MB-453 细胞增殖的影响;流式细胞仪检测细胞周期变化及细胞凋亡;RT-PCR和免疫细胞化学法分别检测Her-2基因mRNA和Her-2、p53蛋白表达水平.结果:VES能抑制乳腺癌MDA-MB-453 细胞增殖,随VES剂量的增加,抑制作用增强.其中20μg/ml VES处理组在48h后受到明显抑制,抑制率达52.25%;经VES处理后细胞出现凋亡,5μg/ml处理48h后,凋亡率为39.52%;药物干预组使肿瘤细胞大量阻滞 于G1期(P<0.01);RT-PCR 和免疫细胞化学法结果表明,VES能抑制Her-2基因mRNA的转录水平,从而抑制其蛋白的表达,也能降低p53蛋白的表达.结论:VES可诱导Her-2、p53 共表达乳腺癌MDA-MB-53细胞凋亡,抑制增殖.其机制可能是通过抑制Her-2基因mRNA及蛋白的表达和通过抑制突变型p53蛋白的表达使细胞停滞于G1有关.%Objective :To investigate the effects of VES inducing human breast cancer cell line MDA - MB - 453 apoptosis , changes of the expression of Her - 2 and p53 before and after apoptosis and changes of the mRNA of Her - 2 gene. Methods : MTT assay was used to detect MDA - MB -453 cell proliferation. Cell cycle and apoptosis were analyzed by flow cytometry. The level of Her -2 mRNA was detected by RT - PCR and the level of Her -2 and p53 expression was also detected by immunofluorescence cytochemicalmethods. Results : The growth of MDA MB - 453 cells was significantly inhibited in a dose - dependent manner after 20mg/L VES treatment. 20μg/ml VES inhihited relatively cells growing by 52. 25％ after treating for 48h. Apoptosis was discovered when treating VES, apoptosis rate was 39. 52％ by 5 μg/ml VES treating for 48h. VES can hinder most of treated group cells in G1 stage and the protein expression of Her -2 and p53 was up regulated after VES treatment. The level of Her - 2 mRNA decreased by using RT - PCR assay and the level of Her - 2 expression was also decreased by using immunofluorescence cytochemical assay after VES treatment. Conclusion : VES could inhihit MDA - MB - 453 cell proliferation and induce apoptosis, the mechanism probably relates to the level of Her - 2 mRNA and Her - 2 expression decreased and inhihit the expression of p53 to cause cell cyecle blocking in G1 stage.
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