缩氨基硫脲-喹唑啉衍生物S-2BEBD对A549细胞增殖的影响及机制研究  

Proliferation inhibition and apoptosis mechanism of thiosemicarbazone-quinazoline derivative S-2BEBD on A549 cells

作　　者：刘海彬[1] 沈继伟 边圣杰 吴英良[2] LIU Hai-bin;SHEN Ji-wei;BIAN Sheng-jie;WU Ying-liang(School of Biomedical&Chemical Engineering,Liaoning Institute of Science and Technology,Benxi Liaoning 117004,China;School of Pharmaceutical Engineering,Shengyang Pharmaceutical University,Shenyang 110016,China)

机构地区：[1]辽宁科技学院生物医药与化学工程学院,辽宁本溪117004 [2]沈阳药科大学生命科学与生物制药学院,辽宁沈阳110016

出　　处：《中国药理学通报》2018年第11期1616-1621,共6页Chinese Pharmacological Bulletin

基　　金：辽宁省博士启动基金项目(No 20170520417);辽宁科技学院引导基金项目(No Yd201606)

摘　　要：目的探究含有缩氨基硫脲结构的喹唑啉化合物S-2BEBD对肺癌A549细胞增殖的抑制作用及诱导凋亡的机制。方法 MTT法检测不同浓度的S-2BEBD和对照药拉帕替尼(LPTN)对肺癌细胞系A549增殖的抑制作用;倒置显微镜和荧光显微镜观察S-2BEBD对A549细胞形态学的影响;流式细胞术(FCM)考察S-2BEBD与LPTN对A549细胞周期的影响;蛋白免疫印迹法检测LPTN和S-2BEBD对EGFR通路,以及下游信号蛋白的影响。结果肺癌细胞系A549经不同浓度LPTN和S-2BEBD作用后,增殖抑制率呈剂量和时间依赖性。形态学检测表明,S-2BEBD以时间依赖性和剂量依赖性的方式抑制A549细胞的增殖。Western blot结果证明,S-2BEBD可明显下调p-EGFR、p-Akt、p-ERK l/2蛋白的表达水平,且呈剂量依赖性,而对非磷酸化的EGFR、Akt、ERK l/2蛋白的表达无明显影响,并且cleaved caspase-3的表达随S-2BEBD浓度增高而增加。同时,LPTN和S-2BEBD通过诱导促凋亡蛋白Bax的激活,抑制抗凋亡蛋白Bcl-2的活性,诱导A549细胞的凋亡。结论缩氨基硫脲-喹唑啉衍生物S-2BEBD对A549细胞的增殖抑制作用呈剂量和时间依赖性,其抗肿瘤的作用机制可能为诱导细胞凋亡。Aim To investigate the effect of quinazoline compound S-2BEBD containing thiosemicarbazone structure on the proliferation and mechanism of apoptosis of human lung cancer A549 cells.Methods MTT assay was used to detect the inhibitory effects of S-2BEBD and LPTN at different concentrations on the proliferation of lung cancer cell line A549.The S-2BEBD influence on A549 cell morphology was observed by inverted microscope and fluorescence microscope.The effect of S-2BEBD and LPTN on cell cycle of A549 cells were examined by flow cytometry(FCM).Western blot was used to detect the effect of LPTN and S-2BEBD on EGFR pathway and downstream signaling proteins.Results The proliferation inhibition rate of lung cancer cell line A549 was dose-and time-dependent after treated with different concentrations of LPTN and S-2BEBD.Morphological analysis showed that S-2BEBD could inhibit the proliferation of A549 cells in a time-and dose-dependent manner.The results of Western blot showed that S-2BEBD could significantly down-regulate the expression levels of p-EGFR,p-Akt,p-ERK 1/2 protein in a dose-dependent manner,but it had no significant effect on the expression of non-phosphorylated protein EGFR,Akt,ERK 1/2.Moreover,the expression of cleaved caspase-3 increased with the increase of S-2BEBD concentration.LPTN and S-2BEBD induced apoptosis of A549 cells by inducing the activation of pro-apoptotic protein Bax and inhibiting the activity of anti-apoptotic protein Bcl-2.Conclusions The inhibitory effect of thiosemicarbazoline-quinazoline derivative S-2BEBD on the proliferation of A549 cells is dose-and time-dependent,and its anti-tumor mechanism may be via inducing apoptosis.

关 键 词：喹唑啉 缩氨基硫脲 A549细胞 抗肿瘤 增殖 凋亡 机制 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]