选择性COX-2抑制剂对非小细胞肺癌细胞株毒性及分子机制的研究

Toxicity of selective COX-2 inhibitor celecoxib on non-small cell lung cancer cell lines and the molecular mechanism in vitro

目的:探讨选择性环氧化酶-2(cyclooxy-genase-2,COX-2)抑制剂Celecoxib对非小细胞肺癌(non-small cell lung cancer,NSCLC)细胞株增殖与凋亡的作用以及相关分子机制。方法:对3种NSCLC细胞株A549(腺癌)、GLC82(腺癌)和SW1573(肺泡细胞癌)使用MTT法测定细胞生长抑制率;Hoechest33258荧光染色观察细胞凋亡;流式细胞仪分析细胞凋亡及细胞周期分布;West-ern blot法检测COX-2、磷酸化AKT(p-AKT)、总丝苏氨酸激酶(serine/threonine kinase,AKT)、磷酸化ERK(p-ERK)及总细胞外信号调节激酶(ex-tracellular signal-regulated kinase,ERK)蛋白的表达。结果:Celecoxib抑制A549、GLC82和SW1573细胞增殖的IC50值分别为14·7、10·6和18·6μmol/L;Celecoxib对3种NSCLC细胞株都有较明显的凋亡诱导作用,Celecoxib作用12h凋亡率增加,24～48h更明显,Celecoxib10～40μmol/L作用可见凋亡率明显增加;Celecoxib作用于GLC82细胞后检测到胞内p-AKT及p-ERK蛋白表达下调。结论:COX-2抑制剂Celecoxib在体外能抑制细胞信号传导AKT及ERK通路的活化,诱导NSCLC细胞发生凋亡,抑制细胞增殖。

OBJECTIVE： To study the effect of growth inhibition and apoptosis induction in non-small cell lung cancer （NSCLC） cell lines by selective COX-2 inhibitor celecoxib and the cell molecular mechanism. METHODS： MTT assay was used to detect the growth inhibition of celecoxib on 3 NSCLC cell lines A549 （adenocarcinoma）, GLC82 （adenocarcinoma）, SW1573 （bronchioalveolar carcinoma）. Hoechest33258 staining was used to detect cell apoptosis. Flow cytometric analysis was used to assess the cell cycle distribution and apoptosis. Western blot analysis was used to detect the expressions of COX-2, p-AKT, AKT, p-ERK and ERK proteins. RESULTS： The IC50 of celecoxib on A549, GLC82 and SW1573 were 14.7, 10.6 and 18.6 μmol/L, respectively. Celecoxib induced apoptosis in NSCLC cell lines. Apoptosis was detected after the treatment for 12 hours and stronger for 24-48 hours. Apoptosis was detected under 10-40 μmol/L concentration celecoxib treatment. P-AKT and p-ERK proteins were detected to be downregulated after the celecoxib treatment. CONCLUSION： Selective COX-2 inhibitor celecoxib can inhibit the cell signal transduction AKT and ERK pathway in NSCLC and induce apoptosis and cell growth inhibition.