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目的:揭示环氧合酶2(COX-2)在肾癌细胞中的表达情况,通过COX-2抑制剂NS398作用肾癌细胞探讨非甾体抗炎药(NSAID)对肾癌细胞增殖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25,50,100,150及200μmol/L的剂量加入细胞中作用24及48h后,MTT法检测NS398对肾癌细胞增殖的影响;作用24h后,流式细胞仪测定细胞凋亡的情况,EIA法测定细胞中前列腺素E(2PGE2)含量的变化,Western blotting测定COX-2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,随着浓度升高,凋亡峰亦越来越增高(P<0.05);NS398可抑制PGE2释放,并且这种抑制作用呈剂量效应,与对照组相比有统计学意义(P<0.05);不同浓度NS398作用下的肾癌786-0细胞中,COX-2的表达明显减弱,且呈剂量梯度下降。结论:NS398通过诱导凋亡来抑制肾癌786-0细胞的增殖;NS398诱导肾癌786-0细胞的凋亡作用机制可能是通过抑制COX-2的表达,降低肾癌786-0细胞PGE2的合成,减少前列腺素对肿瘤细胞增殖的刺激作用来抑制增殖和促进凋亡的。
Objective: To reveal the expression of cyclooxygenase 2 (COX-2) in human renal cell carcinoma (NSCLC) and to explore the effect of non-steroidal anti-inflammatory drug (NSAID) on proliferation of renal cell carcinoma Influence and its possible mechanism of action. Methods: Human renal cell carcinoma 786-0 cells were cultured by standard cell culture method. After NS398 cells were added to the cells at doses of 25, 50, 100, 150 and 200 μmol / L for 24 and 48 h respectively, MTT assay was used to detect the proliferation of renal carcinoma cells . After 24h, the cell apoptosis was measured by flow cytometry. The content of prostaglandin E (2PGE2) in cells was measured by EIA and the expression of COX-2 protein was detected by Western blotting. 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); NS398 cells treated with 786-0 cells Significant sub-diploid apoptotic peak appeared at G0 / G1 phase of cell cycle, and the peak of apoptosis increased with increasing concentration (P <0.05). NS398 could inhibit the release of PGE2 The inhibitory effect was dose-effect, which was statistically significant compared with the control group (P <0.05). The expression of COX-2 in 786-0 cells with different concentrations of NS398 significantly decreased and the dose gradient decreased. Conclusion: NS398 can inhibit the proliferation of 786-0 cells by inducing apoptosis. The mechanism of NS398 inducing apoptosis of 786-0 cells may be through inhibiting the expression of COX-2 and decreasing the PGE2 of 786-0 cells Synthesis, reduce prostaglandin stimulation of tumor cell proliferation to inhibit proliferation and promote apoptosis.