摘要
目的揭示COX-2选择性抑制剂NS398对肾癌细胞的增殖和凋亡作用的影响,及其可能的作用机制。方法采用标准的细胞培养方法对人肾癌786-0细胞进行培养,将NS398分别以不同浓度剂量加入细胞中作用24及48h后,应用流式细胞仪检测凋亡,应用RT-PCR和Western blotting检测COX-2和Bcl-2表达的改变。结果NS398对肾癌786-0细胞具有较强的抑制作用,且这种抑制作用随浓度和时间的增加而增大,呈浓度依赖关系(P<0.05);RT-PCR和Western Blot结果表明,不同浓度NS398作用下的肾癌786-0细胞中,COX-2和Bcl-2的表达明显减弱,且呈剂量梯度下降。NS398作用于肾癌786-0细胞24h后,在细胞周期G0/G1期前出现明显的亚二倍体凋亡峰,随着浓度的升高,凋亡峰亦越来越高(P<0.05)。结论肾癌786-0细胞中存在着COX-2的过表达;选择性COX-2抑制剂NS398通过诱导凋亡来抑制肾癌786-0细胞的增殖,其机制可能是通过抑制COX-2的表达及下调Bcl-2来完成的。
[Objective] To evaluate the effects of NS398, a selective eyelooxygenase-2 （COX-2） inhibitor on cellular proliferation and apoptosis in human RCC cell line, and to examine possible mechanisms responsible for the anti-proliferative effects of NS398. [Methods] Human RCC cell line 786-0 was maintained in culture using standard technique and treated with NS398 at doses of 25, 50, 100, 150 and 200 μM, respectively. Cellular inhibition was measured by MTT at 24 and 48 h. Apoptosis was evaluated by Flow Cytometry. Reverse transeription-polymerase chain reaction （RT-PCR） and Western blotting were used to examine the expression of COX-2 and Bcl-2 at mRNA and protein levels. [Results] NS398 exhibited significant cell growth inhibition at 24 hr and 48 hr in the 786-0 cells in time and concentration-dependent manner. COX-2 and Bcl-2 were significantly decreased by NS398 both at mRNA and protein levels. NS398 inhibited proliferation of 786-0 cells through G0/G1 phase arrest along with induction of apoptosis in dose-dependent manner. [Conclusions] Over-expression of COX-2 mRNA and protein exist in human 786-0 RCC cell line and COX-2 may play a crucial role in carcinogenesis of RCC. NS398 can suppress the proliferation of 786-0 RCC cells by induction of apoptosis. The possible mechanism of induction of apoptosis in 786-0 cells is inhibition of COX-2 expression and suppression of Bcl-2 by NS398. The results suggest that COX-2 may become a new target gene for RCC treatment, and NS398 may be a potentially effective agent against human RCC expressing COX-2.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2008年第9期1153-1157,1161,共6页
China Journal of Modern Medicine
基金
Scientific Research Project for Institute of Higher Education of Education Bureau in Liaoning Province (No.:202013145)
