苯噻啶对人肺腺癌细胞增殖、迁移与侵袭能力的影响  

Effects of pizotifen on proliferation,migration and invasion of human lung adenocarcinoma cells

作　　者：郑江霞 束军[1] 汪新 杨彩梅 沈继龙 Zheng Jiangxia;Shu Jun;Wang Xin;Yang Caimei;Shen Jilong(Dept of Respiratory Medicine,The Fourth Affiliated Hospital of Anhui Medical University,Hefei 230011;The Key Laboratory of Microbiology and Parasitology of Anhui Province,The Key Laboratory of Zoonoses of High Institutions in Anhui,Hefei 230032)

机构地区：[1]安徽医科大学第四附属医院呼吸内科,合肥230011 [2]病原生物学安徽省重点实验室,人畜共患病安徽高校省级重点实验室,合肥230032

出　　处：《安徽医科大学学报》2022年第8期1294-1299,1304,共7页Acta Universitatis Medicinalis Anhui

基　　金：安徽高校自然科学研究重点项目(编号:KJ2018A0204)。

摘　　要：目的观察不同浓度苯噻啶(Pizotifen)对人肺腺癌A549和PC9细胞增殖、迁移与侵袭能力的影响,并初步探究其机制。方法取对数生长期的A549和PC9细胞,分为对照组(细胞对照组为培养基直接作用于细胞,溶剂对照组为含有0.1%DMSO的培养基作用于细胞)和实验组(浓度分别为10、20、30、40μmol/L的苯噻啶作用于细胞)。CCK-8法检测人肺腺癌细胞的增殖能力,划痕实验和Transwell迁移实验检测其迁移能力,Transwell侵袭实验检测其侵袭能力,ELISA法检测Wnt3a/β-catenin信号通路蛋白、上皮-间质转化(EMT)相关蛋白E-钙黏蛋白(E-cad)和基质金属蛋白酶-9(MMP-9)的表达水平。结果与溶剂对照组相比:CCK-8结果显示,实验组A549和PC9细胞活力降低,出现不同程度的增殖抑制现象,在20、30、40μmol/L浓度组有一定的时间依赖性(P<0.05);划痕实验结果表明,实验组划痕愈合率小于对照组,苯噻啶浓度越高,划痕愈合率越低(P<0.05);Transwell迁移和侵袭实验结果显示,实验组穿过小室的肺腺癌细胞数量明显低于对照组,具有浓度依赖性(P<0.05);ELISA结果显示,实验组细胞中Wnt3a、β-catenin、MMP-9蛋白浓度与对照组相比均显著降低,并且随着苯噻啶浓度增加,Wnt3a、β-catenin、MMP-9蛋白浓度逐渐降低(P<0.05);实验组细胞中E-cad蛋白浓度与对照组相比显著增高,并且随着苯噻啶浓度增加,E-cad浓度逐渐增高(P<0.05)。结论在体外细胞实验中,苯噻啶对人肺腺癌A549、PC9细胞的增殖、迁移和侵袭能力具有抑制作用,抑制作用的强弱依赖于苯噻啶的浓度和作用时间。其作用机制,可能包括Wnt3a/β-catenin-EMT通路的阻断。Objective To investigate the effects of Pizotifen on proliferation,migration and invasion of A549 and PC9 cells of human lung adenocarcinoma and its possible mechanisms.Methods The logarithmic phase of A549 and PC9 cells were used for experiments,which were divided into control groups(cell control group and DMSO solvent control group)and experimental groups(including 10,20,30 and 40μmol/L Pizotifen,respectively).CCK-8 assay was used to evaluate the effects of Pizotifen on A549 and PC9 cells proliferation.Wound-healing assay andTranswell assay were then performed to investigate whether Pizotifen affected the migration and invasion properties of A549 and PC9 cells.Finally,ELISA was used to determine the expression levels of Wnt3a/β-catenin signaling pathway related proteins and epithelial-mesenchymal transition(EMT)related proteins such as E-cadherin(E-cad)and matrix metalloproteinase-9(MMP-9)in A549 and PC9 cells after Pizotifen treatment.Results Compared with the solvent control group,the assay of CCK-8 in experimental groups showed that the cell vitality gradually decreased after Pizotifen treatment for 24 h and 48 h,and the inhibition rate of cell vitality was directly proportional to the action time of Pizotifen(P<0.05).At the same treatment time,the cell viability of A549 and PC9 in the experimental groups decreased with the increase of Pizotifen concentrations(20,30 and 40μmol/L).The test of Wound-healing showed that Pizotifen had an inhibitory effect on the migration of A549 and PC9 cells,and the higher the concentration of Pizotifen was,the lower the cell migration rate was(P<0.05).The experiment of Transwell showed that Pizotifen had an inhibitory effect on the migration and invasion of A549 and PC9 cells in a concentration-dependent manner(P<0.05).ELISA showed a decrease in the level of Wnt3a,β-catenin and MMP-9 protein and an increase in E-cad protein in the experimental groups compared with the solvent control group.Conclusion Pizotifen may inhibit the proliferation,migration and invasion of human

关 键 词：苯噻啶 人肺腺癌 信号通路 上皮-间质转化 

分 类 号：R734.2[医药卫生—肿瘤]