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目的观察COX-2选择性抑制剂NS398的对肾癌细胞增殖和凋亡作用的影响及其作用机制。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25、50、100、150、200μmoL/L的剂量加入细胞中作用24及48 h后,应用MTT法检测NS398对肾癌细胞增殖的影响;将NS398以50、100及200μmol/L的浓度作用于肾癌786-0细胞24 h后,用流式细胞仪测定细胞凋亡的情况;将NS398分别以25、50、100、150、200μmol/L的剂量加入细胞中作用24 h后,应用RT-PCR和Western blotting分别检测肾癌786-0细胞中COX-2和bcl-2的mRNA及其蛋白表达的情况。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);NS398作用肾癌786-0细胞24 h后,在细胞周期G0/ G1期前出现明显的亚二倍体凋亡峰,随着浓度升高凋亡峰亦越来越增高(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无论在mRNA水平还是在蛋白水平均明显减弱,且呈剂量梯度下降。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,导致bcl-2抗凋亡活性的降低来完成的。
Objective To observe the effect of COX-2 selective inhibitor NS398 on proliferation and apoptosis of renal cell carcinoma and its mechanism. Methods Human renal cell carcinoma 786-0 cells were cultured by standard cell culture method. After NS398 cells were treated with 25, 50, 100, 150 and 200 μmol / L respectively for 24 and 48 h, MTT assay was used to detect NS398 On the proliferation of renal cell carcinoma; NS398 at 50, 100 and 200μmol / L concentration of renal cell carcinoma 786-0 cells for 24 h, the apoptosis was measured by flow cytometry; the NS398 were 25, 50, 100, 150 and 200μmol / L were added into the cells for 24 h, RT-PCR and Western blotting were used to detect the mRNA and protein expression of COX-2 and bcl-2 in renal cell carcinoma 786-0 cells . Results NS398 had a strong inhibitory effect on 786-0 human renal cell carcinoma cells, and this inhibitory effect increased with concentration and time increasing in a concentration-dependent manner (P <0.05) h, there was a significant sub-diploid apoptotic peak before the G0 / G1 phase of the cell cycle, and the apoptotic peak was also increased with increasing concentration (P <0.05). The results of RT-PCR and Western Blot showed that, The expression of COX-2 and bcl-2 in 786-0 cells with different concentrations of NS398 significantly decreased both at the mRNA and protein levels, with a dose-dependent decrease. Conclusion The overexpression of COX-2 exists in 786-0 cell line of renal cell carcinoma, and NS398, a selective COX-2 inhibitor, inhibits the proliferation of 786-0 cell line by inducing apoptosis. The mechanism may be that COX-2 Expression, leading to a decrease in the anti-apoptotic activity of bcl-2.