塞来昔布对人肺腺癌细胞株A549的放射增敏效应及细胞迁移力的影响

Radiosensitization of celecoxib on human lung adenocarcinoma cell line A549 and inhibition of migration ability in vitro

目的 观察选择性COX-2抑制剂塞来昔布（celecoxib）对人肺腺癌细胞株A549的放射增敏效应及抑制细胞迁移力的作用.方法 选用人肺腺癌细胞株A549作为研究对象.选择适当浓度的塞来昔布,设置对照组、药物组、单纯放射组和放射加药组,成克隆分析法测定塞来昔布对细胞的放射增敏效应,细胞划痕试验观察A549细胞的迁移力,酶联免疫吸附实验（ELISA）检测细胞培养上清液中基质金属蛋白酶（MMP）-2的含量.结果 细胞划痕试验结果显示,放射加药组比单纯放射组和药物组的无细胞划痕区均增宽,细胞迁移力均明显下降,且随着药物浓度的增加,抑制细胞迁移的能力增强.30和50 μmol/L塞来昔布对A549细胞显示出浓度依赖性放射增敏作用,放射加药与单纯放射组相比,SF2、D0、Dq出现不同程度的下调.ELISA检测发现,细胞培养上清液中MMP-2的含量,药物组较对照组,放射加药组较单纯放射组均明显下降（t=3.78、5.79,P＜0.05）,但单纯放射组和对照组对细胞分泌MMP-2的抑制效应不明显（t=2.73、2.38,P＞0.05）.结论 塞来昔布在体外试验中对肺腺癌细胞株A549具有浓度依赖性放射敏感性,机制可能为降低了细胞对放射的亚致死损伤修复能力.塞来昔布可能通过抑制A549细胞分泌MMP-2,从而抑制细胞的迁移能力.

Objective To investigate the effects of radiosensitivity enhancement and inhibition of migration ability of human lung adenocareinoma cells by celecoxib, a selective cyclooxygenase （ COX）-2 inhibitor. Methods Human lung adenocarcinoma cells of the line A549 were cultured and then inoculated into six-well plates and randomly divided into 4 groups： control group, eelecoxib group administered with celecoxib at the subtoxic doses 30 and 50 la, mol/L, irradiated group exposed to 0, 1, 2, 4, 6, or 8 Gy by linear accelerator, and combined treatment （celecoxib ＋ irradiation） group. The radiosensitizing effect of celecoxib was assessed by c]onogenie cell survival test. The migration ability of the A549 ceils was measured by scratch-wound test and the content of metalloproteinase-2 （MMP-2） in culture supernatant was detected with ELISA. Results The sensitization enhancement ratio of the eelexib group was increased dose- dependently. The values of DO , Dq, SF2 and D0.01 of the celecoxib ＋ irradiation group were all significantly lower than those of the irradiated group. Scratch-wound test showed that the no-scratch area of the celecoxib ＋ irradiation group and celecoxib group were all significantly wider than those of the mere irradiation and control groups and there was a dose-dependent manner, and the no-scratch area of the celecoxib ＋ irradiation group was wlider than that of the eelecoxib group. ELISA showed that the MMP-2 levels in the supernatant of the celecoxib group and celecoxib ＋ irradiation group were respectively significantly lower than those of the control group and mere irradiated group （t = 3.78, 5. 79,3. 15, P 〈 0.05）, however, there was not significant difference between the mere irradiation and control groups （ t = 2. 73, 2. 38, P 〉 0. 05 ）. Conclusions Celecoxib enhances concentration-dependently the radiosensitivity of human lung carcinoma cell and inhibits the secretion of MMP-2 of the carcinoma cells, thus inhibiting their migration ability.