摘要
目的探讨环氧化酶-2(COX-2)抑制剂NS398对肾癌细胞786-O增殖和凋亡的影响及其机制。方法同浓度NS398作用体外培养786-O细胞后,采用噻唑蓝(MTT)法检测24、48、72、96h后肾癌细胞786-O的增殖活性。30、180μmol/L的NS398作用786-O细胞72h后,RT-PCR法检测COX-2 mRNA表达;免疫细胞化学检测COX-2蛋白表达;酶联免疫吸附测定法(ELISA)检测前列腺素E2(PGE2)释放水平;流式细胞仪检测细胞凋亡情况。结果NS398可以抑制786-O细胞的增殖,呈时间和剂量依赖型。RT-PCR法和免疫细胞化学检测显示NS398作用后能降低COX-2 mRNA和蛋白表达。ELISA检测显示NS398作用后PGE2释放水平呈下调趋势;流式细胞仪检测表明,30、180μmol/L NS398处理组凋亡率分别为(14.3±1.4)%、(31.5±2.1)%,与对照组凋亡率(2.1±0.4)%比较,凋亡率显著上升(P<0.05)。结论NS398可能通过COX-2依赖性途径抑制肾癌细胞增殖和诱导其凋亡。
Objective To study the proliferative and apoptotic effects of NS398,a selective COX-2 inhibitor,on renal carcinoma cell line 786-O and its possible mechanisms. Methods 786-O cells were treated with different concentrations of NS398 for 24h, 48h,72h and 96h. The proliferation of 786-O cells was studied by MTT assay. The expression of COX-2 mRNA was analyzed by RT-PCR. Immuno cytochemistry was used to detect the expressions of COX-2. The release levels of PGE2 was measured by enzymelinked immunosorbent assay(ELISA). Flow cytometric analysis was used to detect the apoptosis status of 786-O cells. Results NS398 resulted in significant antiproliferative effects on 786-O cells in dose and timedependent. RT-PCR showed the expressions of COX-2 were significantly inhibited by NS398. Immunocytochemistry showed the expressions of COX-2 were significantly inhibited by NS398. The release levels of PGE2 were decreased compared with those of the control group. Apoptosis rate was (31.5±2. 1) and (14.3±1.4) % in the 786-0 cells treated with NS398 in different concentrations of 180μmol/L and 30μmol/L, which was sig- nificantly higher than that in the control group (2.1±0.4)%. Conclusion NS398 can inhibit proliferation of renal carcinoma cell line 786-O and induce its apoptosis in vitro,which may contributed to the COX-2 dependent pathway.
出处
《重庆医学》
CAS
CSCD
2008年第14期1525-1527,1530,共4页
Chongqing medicine