西乐葆和雷帕霉素联合作用对人乳腺癌MDA-MB-231细胞的抗肿瘤效应

Antitumor efficacy of combination of celecoxib and rapamycin in MDA-MB-231 human breast cancer cells

目的观察西乐葆和雷帕霉素联合作用对于人乳腺癌MDA-MB-231细胞的抗肿瘤效应。方法不同浓度的西乐葆（20、40、60、80μmol／L）、雷帕霉素（1、10、100、1000nmol／L）单独及60μmol／L西乐葆和100nmol／L雷帕霉素两者联合作用对MDA-MB-231细胞生长的影响通过噻唑蓝（MTT）比色法检测。对细胞周期和凋亡的影响使用流式细胞仪进行分析。蛋白印迹实验检测HER2和HER3的表达情况及磷酸化Akt（473）水平。结果西乐葆、雷帕霉素对于MDA-MB-231细胞生长抑制具有浓度和时间依赖性。60μmol／L西乐葆和100nmol／L雷帕霉素联合应用细胞生长抑制率及凋亡率为[（88．0±8．0）％，（32．5±3．0）％]，与两者单独应用细胞生长抑制率及凋亡率[（52．0±5．0）％，（12．6±2．0）％；（54．0±6．0）％，（7．2±2．0）％]比较差异有统计学意义（P〈0．01）。联合应用对MDA-MB-231细胞抗肿瘤效应与降低细胞HER2和HER3的表达及磷酸化Akt（473）水平有关。结论西乐葆和雷帕霉素联合能增强对人乳腺癌DA-MB-231细胞的抗肿瘤效应，对于HER2、HER3阳性乳腺癌具有潜在的临床应用价值。

Objective To explore the antitumor efficacy of combination of celecoxib and rapamycin in MDA-MB-231 human breast cancer cells. Methods The effects of different concentrations of celecoxib （ 20,40,60 and 80 μmol/L） alone, rapamycin （ 1,10,100,1000 nmol/L） alone, or 60 μmol/L celecoxib in combination with 100 nmol/L rapamycin on cell growth were detected by using MTT. The cell cycle and apoptosis were analyzed by flow cytometry. The expression of HER2 and HER3 and level of phosphorylation of Akt （473） were also detected by using method of Western blot. Results Celecoxib alone or rapamycin alone markedly inhibited MDA-MB-231 cell growth at a dose- and time-dependent manner. Compared with 60 μmol/L celecoxib or 100 nmol/L rapamycin alone treatment ,60 μmol/L celecoxib in combination with 100 nmol/L rapamycin produced synergistic inhibitory effects on cell growth and apoptosis [ （88.0±8.0）% vs （52.0±5.0）% or （54.0±6.0）%,（32.5±3.0）% vs （12.6±2.0）% or （7.2±2.0） % ] （ P 〈 0.01 ）. Tile enhanced antitumor effect of the combined agents was associated with the decreased expression of HER2, HER3 and level of phosphorylation of Akt （473） in MDA-MB-231 cells. Conclusion Enhanced anti-cancer effect is achieved in MDA-MB-231 human breast cancer cells by combining celecoxib and rapamycin. The combination of celecoxib and rapamycin may increase the clinical potential benefits of these agents to sub-populations of HER2 and HER3-positive breast cancer.