摘要
目的曲马多作用于肺癌细胞是否会影响肿瘤细胞的生长和转移,目前国内外报道较少。文中旨在探讨曲马多对人肺腺癌细胞A549增殖、凋亡和细胞周期的影响。方法人肺腺癌细胞A549培养板接种培养24 h后,分为4组:对照组(等渗盐水)、3μg/m L曲马多组、30μg/m L曲马多组、300μg/m L曲马多组。孵育48 h,采用CCK8法检测细胞增殖能力,Annexin-V FITC/PI双染法检测细胞凋亡,免疫印迹法检测X连锁凋亡抑制蛋白(XIAP)、Survivin、Bcl-2、Bax、Caspase-3蛋白表达水平,流式细胞仪进行肿瘤细胞周期检测。结果与对照组比较,3μg/m L曲马多组、30μg/m L曲马多组、300μg/m L曲马多组吸光度值逐渐降低(P<0.05),且能显著促进细胞的凋亡。与对照组细胞的凋亡率[(2.1±0.3)%]比较,3μg/m L曲马多组[(4.9±0.7)%]、30μg/m L曲马多组[(9.4±1.4)%]、300μg/m L曲马多组[(19.2±2.4)%]逐渐升高(P<0.05)。300μg/m L曲马多组Survivin蛋白、XIAP蛋白相对表达水平[(0.31±0.02)、(0.38±0.04)]较对照组[(0.98±0.04)、(0.99±0.02)]下调,Caspase-3蛋白(1.06±0.03)较对照组(0.97±0.03)上调(P<0.05)。与对照组细胞周期比例比较,3μg/m L曲马多组、30μg/m L曲马多组、300μg/m L曲马多组均能使G0/G1期细胞比例降低,S期及G2/M期比例增加,且具有剂量依赖性(P<0.05)。结论曲马多可抑制人肺腺癌细胞A549的增殖,促进凋亡,细胞周期阻滞;其机制可能与下调Survivin,XIAP蛋白的表达和促进Caspase-3蛋白活化有关。
Objective Few reports are seen domestically on the effect of tramadol(TMD)on the growth and metastasis of lung cancer cells.This study was to investigate the effect of TMD on the proliferation,apoptosis and cell cycle of human lung adenocarcinoma(LAC)A549 cells.Methods We treated human LAC A549 cells with isotonic physiological saline(control)and TMD at 3μg/mL(low-dose TMD),30μg/mL(medium-dose TMD)and 300μg/mL(high-dose TMD),respectively.After 48 hours of intervention,we detected the proliferation of the cells by CCK-8 assay,determined their apoptosis by Annexin-V FITC/PI double staining,measured the protein expressions of XIAP,Survivin,Bcl-2,Bax,and Caspase-3 in the cells by Western blot,and examined the cell cycle by FCM.Results Compared with the control group,tramadol treatment significantly reduced the absorbance value of the LAC A549 cells(P<0.05)and elevated the level of their apoptosis in the low-,medium-and high-dose TMD groups([2.1±0.3]%vs[4.9±0.7]%,[9.4±1.4]%and[19.2±2.4]%,P<0.05),both in a dose-dependent manner.In comparison with the saline control,high-dose TMD group showed markedly down-regulated expressions of Survivin(0.98±0.04 vs 0.31±0.02,P<0.05)and XIAP(0.99±0.02 vs 0.38±0.04,P<0.05)but up-regulated expression of Caspase-3(0.97±0.03 vs 1.06±0.03,P<0.05)in the LAC A549 cells.The ratio of G0/G1 cells was remarkably decreased while those of S and G2/M cells significantly increased,all in a dose-dependent manner(P<0.05).Conclusion Tramadol can inhibit the proliferation,promote the apoptosis and block the cycle of human LAC A549 cells,which may be associated with its abilities of down-regulating the expressions of Survivin and XIAP and promoting the activation of Caspase-3 in the cells.
作者
许彦劼
夏明
XU Yan-jie;XIA Ming(Department of Clinical Medicine,Xuzhou Medical University,Xuzhou 221004,Jiangsu,China;Key Laboratory of Anesthesiology of Jiangsu Province,Xuzhou Medical University/Department of Anesthesiology,Affiliated Hospital of Xuzhou Medical University,Xuzhou 221004,Jiangsu,China)
出处
《医学研究生学报》
CAS
北大核心
2018年第9期927-930,共4页
Journal of Medical Postgraduates
基金
中国博士后基金(2017M611914)
江苏省博士后基金(1701044C)
江苏高校品牌专业建设工程一期项目(PPZY2015B161)
关键词
曲马多
肺腺癌
增殖
凋亡
细胞周期
tramadol
lung cancer
proliferation
apoptosis
cell cycle
