尼美舒利促进下咽癌细胞凋亡并抑制其转移的实验研究

Nimesulide induced apoptosis and inhibited metastasis of hypopharyngeal carcinoma cells

目的 研究尼美舒利[选择性环氧合酶-2（cyclooxygenase-2,Cox-2）抑制剂]对下咽癌细胞系FaDu凋亡和转移能力的影响.方法 应用梯度浓度的尼美舒利（0、10、50、100、500和1000 μmol/L）作用于FaDu细胞,四甲基偶氮唑蓝法（MTT）检测FaDu细胞的相对存活率.在有效药物浓度500μmol/L尼美舒利作用12 h后,吖啶橙染色检测细胞的形态变化.流式细胞术检测FaDu细胞的凋亡率.应用Transwell小室检测尼美舒利对FaDu细胞的侵袭和转移能力的影响.Western blot和反转录-聚合酶链反应（RT-PCR）检测Cox-2、基质金属蛋白酶-9（matrix metalloproteinase,MMP-9）和Caspase-3用药前后的表达变化.结果 MTT结果显示,尼美舒利可以显著降低FaDu细胞相对存活率,其效应具有时间和浓度依赖性（P值均〈0.05）.吖啶橙染色可见细胞凋亡的形态学改变,流式细胞术检测到尼美舒利作用6 h后,FaDu细胞的凋亡率为（32.4±6.1）%（（-x）±s）.尼美舒利使FaDu细胞的侵袭和转移能力下降.Western blot和RT-PCR检测Cox-2、MMP-9表达降低,而Caspase-3的表达升高.结论 选择性Cox-2抑制剂尼美舒利能够抑制FaDu细胞的生长、降低其侵袭和转移能力,促进细胞凋亡.

Objective To investigate the effects of Nimesulide, a selective Cox-2 inhibitor, on the apoptosis, invasion and migration of hypopharyngeal carcinoma cell line （FaDu）. Methods Viabilities of FaDu cells treated with various concentrations of Nimesulide were detected by MTT assay. Morphological changes were observed by acridine orange cytochemistry staining. Apoptosis was examined by flow cytometry.The ability of invasion and migration of cells was detected by Transwell chambers. The mRNA and protein expressions of Cox-2, MMP-9 and caspase-3 in response to Nimesulide were examined by RT-PCR and Western blot, respectively. Results MTT assay showed that the cell surviving rates significantly decreased in time- and concentration-dependent manners （ P 〈 0.05 ）. The typical morphological changes of apoptotic cells were observed. Percent of apoptosis after 6 h Nimesulide treatment was （ 32.4 ± 6.1 ）%. The metastatic cells were decreased by Nimesulide to about 20%. Nimesulide decreased the expressions of Cox-2 and MMP-9, whereas increased expression the of Caspase-3. Conclusion Nimesulide could induce the apoptosis and inhibit metastasis of FaDu cells.