N-(3-三氟甲基苯基)维甲酰胺对人肺腺癌A549细胞增殖及凋亡的影响

Effects of N-(3-trifluoromethyl-phenyl)-retinamide on proliferation and apoptosis of human pulmonary adenocarcinoma A549 cells

目的研究新合成的全反式维甲酸(ATRA)衍生物[N-(3-三氟甲基苯基)维甲酰胺]对人肺腺癌A549细胞株体外增殖及凋亡的影响。方法3种浓度的ATRA衍生物和ATRA(1、5、10mg/L)分别处理A549细胞1、2、3、7d,应用MTT法和流式细胞技术分别观察该ATRA衍生物对A549细胞增殖、凋亡和细胞周期的影响;Western blot分析凋亡相关蛋白表达的变化。结果ATRA衍生物处理组A549细胞增殖抑制率随药物浓度的增加而提高,分别为9.1%、17.8%、47.0%;抑制率亦随药物作用时间的延长而提高,分别为18.1%、20.6%、40.4%、83.4%。10mg/LATRA衍生物可明显诱导A549细胞凋亡,凋亡率达到40.9%,G0～G1期细胞比例明显下降,细胞凋亡峰较对照组明显提前,凋亡过程伴随Bcl-2表达下降,而Bax、Bak、酶原形式的Caspase-3、p53、NF-κB的表达无明显变化。结论ATRA衍生物对A549细胞的增殖抑制作用呈浓度和时间依赖性;并诱导细胞凋亡,可能与Bcl-2蛋白的表达下调有关。

Objective To study the effects of ATRA derivant N-（3-trifluoromethyl-phenyl）-retinamide on proliferation and apoptosis of human pulmonary adenocarcinoma cell line A549 in vitro and its mechanisms. Methods The A549 cells were respectively treated by ATRA and ATRA derivant at the dose of （1,5,10） mg/L for 1,2,3,7 d. The proliferation of A549 cells was observed by MTT assay, and the cell cycle and apoptosis were detected by flow cytometry. The related protein expression was analyzed by Western blot. Results In ATRA-derivant group, the inhibition ratio of A549 cells increased with the increase of ATRA-derivant concentration, 9. 1%, 17. 8% ,47. 0% respectively, also the inhibition ratio increased with the increase of action time, 18.1% ,20.6% ,40.4% ,83.4% respectively. 10 mg/L ATRA derivant induced apoptosis obviously, apoptosis rate was 40. 9% , the G0 -G1 period cell rate decreased obviously, apoptotic peak advanced markedly, with the expression of Bcl-2 decreased without marked change of Bax,Bak, Caspase-3 proenzyme, p53, NF-KB expression. Conclusion ATRA derivant could inhibit the A549 cells proliferation in dose and time-dependent manner, and induce the A549 cells apoptosis by downregulating the expression of Bcl-2 protein.