As_2O_3对ACC-M细胞增殖及VEGF和bFGF表达的影响

Effect of As_2O_3 on Gene Expression of VEGF and bFGF in ACC-M Cells

目的:研究三氧化二砷(As2O3)对人类腺样囊性癌ACC-M细胞体外增殖及表达VEGF和bFGF的影响。探讨As2O3对ACC-M细胞表达VEGF和bFGF的影响。方法:倒置相差显微镜下观察ACC-M细胞生长情况,采用MTT法检测不同浓度As2O3作用不同时间对ACC-M的抑制效应;以RT-PCR、Western blot方法检测As2O3作用后ACC-M细胞的两种血管生成相关因子VEGF和bFGF表达的变化。结果:As2O3抑制ACC-M细胞呈时间-剂量依赖关系。RT-PCR检测显示,As2O3作用后ACC-M细胞VEGF和bFGF基因表达无明显变化。Western blot检测显示,As2O3明显抑制ACC-M细胞VEGF和bFGF蛋白表达,呈时间-剂量依赖关系。结论:As2O3在体外明显抑制ACC-M细胞;As2O3明显抑制ACC-M细胞VEGF和bFGF基因的蛋白产物,其作用机制可能为抗腺样囊性癌的血管形成。

Objective： The aim of this study was to investigate the proliferation effects of As2O3 on salivary adenoid cystic carcinoma-M （ACC-M） cells in vitro and gene expression of VEGF and bFGF within those cells. The possible mechanism of As2O3 on gene expression of VEGF and bFGF was also elucidated. Methods： ACC-M ceils were treated with different concentration of As2O3 for different time. Cytomorphology of ACC-M cells was observed on phasecontrast microscope. Proliferation or suppression effect on cells＇ viability was detected by MTT method. Gene and protein expression of VEGF and bFGF were investigated by RT-PCR and Western blot analysis respectively. Results： Cell viability after As2O3 treatment was markedly suppressed and exhibited as a dose- and time-dependent pattern. RT-PCR revealed no statistically difference between gene expression of VEGF and bFGF. Through Western blot analysis, a negative correlation between As2O3 concentration and amount of protein product of these two cytokines was determined. Conclusions： As2O3 might markedly suppressed ACC-M cells＇ viability with a time-dose reliable pattern in vitro. As2O3 decreases protein product of VEGF and bFGF apparently. Therefore, the agent might suppress vessel formation of ACC-M cells.