摘要
目的:观察COX-2抑制剂塞来昔布对人肝癌细胞株SMMC-7721增殖抑制和凋亡诱导作用,并探讨该作用与ERK1/2信号通路之间的关系。方法:用不同浓度的塞来昔布溶液(0、25、50、75和100μmol/L)处理对数生长期的肝癌SMMC-7721细胞,MTT比色法检测不同时间点各药物浓度组细胞增殖情况;流式细胞术分析不同浓度的塞来昔布溶液对SMMC-7721细胞凋亡的影响;RT-PCR检测COX-2、VEGF和ERK1/2mRNA表达的变化;Western blot检测COX-2、VEGF和ERK1/2蛋白表达的变化。结果:塞来昔布对SMMC-7721细胞增殖具有抑制作用,该作用随着药物浓度的增加和作用时间的延长而增强;流式细胞术检测结果显示出明显的细胞凋亡,且凋亡率随着塞来昔布浓度的增高而升高;随着塞来昔布剂量的增加,COX-2、VEGF和ERK1/2mRNA及蛋白的表达水平逐渐降低。结论:塞来昔布可抑制肝癌SMMC-7721细胞增殖、促使瘤细胞凋亡,且作用呈时间和剂量依赖性,其机制可能与下调ERK1/2信号通路及抑制新生血管生成有关。
Objective:To explore the effects of Celeeoxib on the proliferation and apoptosis of hepatocellular carcinoma cell line SMMC-7721 Cells and elucidated the correlation between the effects of Celecoxib and ERK1/2 pathway. Methods : Cultured SMMC-7721 Cells were treated with different concentration of Celecoxib (0,25,50,75, and 100μmol/L) for the different times,MTT assay was used to test the effect of Celecoxib on the cell proliferation; The rates of apoptosis of cells of treatment with celecoxib were assessed by the flow cytometry;Quantitive real time RT-PCR and western blotting were emplored to examine the expression of COX-2, VEGF and ERK1/2; in Cultured SMMC-7721 cells. Results: Celecoxib could inhibit , in a dose and time dependent manner, the proliferation of SMMC-7721 can signficantly;compared to control ,the apoptotic ceils could be obviously observed and the apoptosis rate significantly increased with increases in concentrations of celecoxib~ Furthermore, The level of COX-2, VEGF, ERK1/2 mRNA and protein expression were significantly down-regulated with increases in concentrations of celecoxib(P〈0.05). Conclusion:The inhibition of proliferation and apoptosis in SMMC-7721 cells can be induced signif- icantly by celecoxib in a dose-and time-dependent manner and its mechanism may be the down regulation of the expression of ERK1/2 and inhibition angiogenesis.
出处
《陕西医学杂志》
CAS
2013年第6期649-651,666,共4页
Shaanxi Medical Journal