鱼油组分共轭亚油酸抑制肺腺癌干细胞成瘤的体内研究  

CLA inhibits proliferation of lung adenocarcinoma stem cells in vivo

作　　者：胡振东[1] 尹荣[1] 许林[1] 李宁[2] 

机构地区：[1]江苏省肿瘤医院胸外科,江苏南京210009 [2]南京军区南京总医院普通外科,江苏南京210002

出　　处：《肠外与肠内营养》2014年第5期301-304,308,共5页Parenteral & Enteral Nutrition

基　　金：南京市科技发展计划项目(201104025)

摘　　要：目的:建立肺腺癌干细胞荷瘤动物模型,体内验证鱼油组分共轭亚油酸(CLA)抑制肺腺癌增殖及转移的作用和分子机制。方法:采用新霉素和克隆挑选技术分别建立稳定表达绿色荧光蛋白(GFP)的A549和SPC-A-1细胞株,体外培养富集干细胞微球,建立肺癌干细胞原位荷瘤裸鼠模型,腹腔注射给予不同组分c9t11-CLA、t10c12-CLA和混合组,定期检测裸鼠体重和肿瘤质量,研究终点采用小动物活体荧光成像技术分别观察肿瘤的转移情况。采用Western blotting检测肿瘤组织COX-2、CXCR4的蛋白表达水平。结果:实验终点各组原发肿瘤重量,A549混合治疗组比各单药治疗组肿瘤质量显著降低(P<0.001),而各CLA单药治疗组比,A549 control组肿瘤质量显著降低(P<0.01),与SPC-A-1组结果相似。各组肿瘤转移情况:在动物活体荧光成像系统下,A549CSC和SPC-A-1 CSC空白对照组出现了全身多处转移;经c9,t11-CLA、t10,c12-CLA治疗显著减少转移位置和比率;而混合治疗组减少尤为明显。Western blotting检测OCT4、COX-2、CXCR4蛋白表达水平。在A549肺腺癌干细胞组,c9t11-CLA、t10c12-CLA均不同程度地抑制OCT4、COX-2、CXCR4蛋白,混合治疗组抑制作用最强。在SPC-A-1肺腺癌干细胞组结果类似。结论:c9,t11-CLA、t10,c12-CLA及其混合物能明显抑制肺腺癌干细胞荷瘤裸鼠原发肿瘤和转移灶的生长;CLA组分能明显抑制肿瘤组织OCT4、COX-2和CXCR4蛋白的表达。Objective：This study was aimed to establish xenografts of lung adenocarcinoma stem cell in nude mice,validate the inhibitory effect of CLA on proliferation and migration in vivo,and investigate the underlying molecular mechanisms.Methods：By neomycin and clone selection,GFP stably expressed A549 and SPC-A-1 cell lines were established and then tumorosphere of these 2 cell lines were cultured and collected.The tumorosphere were then disassociated and injected injected into nude mice subcutaneously to establish xenografts.The nude mice were divided into four groups：blank control,c9t1 1-CLA group,t10c12-CLA group,and combined group （c9t1 1-CLA ＋ t10c12-CLA）.The CLA was injected intraperitoneally and normal saline were used in blank control.Weight of the nude mice and xenografts were recorded and metastasis was observed by Small animals in vivo fluorescence imaging system.Western blot was applied to examine the protein level of COX-2,CXCR4.Results：As for weights of A549 xenografts,the combined therapy significantly decreased the tumor weight compared with the two monotherapy groups （P ＜0.01）,and both the two CLA monotherapy reduced tumor weight compared with the blank control （P ＜ 0.01）,while there was no difference between 2 monotherapy groups.Similar results were observed for SPC-A-1 xenografts.For metastasis,by in vivo fluorescence imaging system,systemically multiple metastasis was observed in A549 and SPC-A-1 control groups,while metastasis sites and ratio were significantly reduced in the CLA monotherapy groups or combined groups,which was most significant in the combined group.Western blot suggested that the protein level of OCT4,COX-2,and CXCR4 were reduced by CLA treatment,and the inhibitory effect was strongest in the combined group.Conclusion：c9t1 1-CLA and t10c12-CLA,either alone or in combination,could significantly inhibit proliferation of lung adenocarcinoma xenografts,and reduced protein level of OCT4,COX-2,and CXCR4 in vivo.

关 键 词：鱼油组分共轭亚油酸 肺腺癌干细胞 表面标记物 增殖 迁移 荷瘤裸鼠模型 

分 类 号：R734.2[医药卫生—肿瘤]