摘要
目的研究人乳腺癌斑马鱼移植瘤模型的建立方法及其相关的蛋白表达。方法分别选用MCF-7、T-47D、MDA-MB-231细胞系显微注射至斑马鱼幼鱼体内,考察不同乳腺癌细胞系在斑马鱼体内的定植迁移、细胞数量变化、细胞分布以及对斑马鱼肠下静脉丛(subintestinal veins,SIVs)的影响,建立乳腺癌斑马鱼移植瘤模型;并考察JAK抑制剂对斑马鱼移植肿瘤的影响,并对相关蛋白表达的情况进行蛋白印迹分析。结果在接种细胞数大于每只200时,斑马鱼的肿瘤模型成功率大于比例0.90;MB-231移植瘤在斑马鱼体内具有明显的迁移特征,这种特征可被JAK抑制剂tofacitinib抑制(P<0.01);同时tofacitinib可明显减少移植瘤模型体内MB-231的数量(P<0.01);移植瘤可促进SIVs增生,酪氨酸抑制剂PTK787可明显地抑制移植瘤引起的SIVs的生长(P<0.01);蛋白印迹结果显示,4 d斑马鱼胚胎自身HER2有表达,T-47D斑马鱼模型组组织中HER2表达增加;MB-231斑马鱼模型组组织中VEGFa高表达;tofacitinib处理后的移植瘤其p53表达增强。结论人源肿瘤通过微量显微注射方法可成功建立斑马鱼乳腺癌移植瘤模型,该模型可用于抗肿瘤药物的初步筛选研究。
Aim To investigate the modeling of breast cancer in zebrafish embryos and its related protein expression. Methods48 hpf wild type AB / TG( Transgenic) zebrafishs were micro-injected with breast cancer cell line: MCF-7,T-47 D,MDA-MB-231 respectively,the relationship between the number of tumor and model application was investigated,and the number of subintestinal veins( SIVs) was detected under confocal microscope,as well as the metastasis of tumor cells in embryos; then the zebrafish xenografts of MB-231 were co-cultured with tofacitinib /ptk787 for 48 h,optical density( OD) of the cell survival and subintestinal veins( SIVs) were evaluated under confocal microscope,and Western blot( WB) analysis was used to test microcircumstances related protein. Results When the number of inoculated cells was more than 200 per embryo,xenograft model rate woule be more than 0. 90; MB-231 xenografts showed metas-tasis feature in zebrafish,which could be inhibited by tofacitinib( P < 0. 01),while the number of xenograft MB-231 cells was reduced significantly( P < 0. 01); in another zebrafish xenografts SIVs assay,the tumor could promote the proliferation of SIVs,and 4 mg · L- 1PTK787 showed inhibiton effect( P < 0. 01).Western blot showed 4d T-47 D xenograft zebrafish got more HER2 expression than AB embryos; VEGFa expression in zebrafish MB-231 model group was higher,and model zebrafish P53 expressi was higher after treated by tofacitinib. Conclusion A zebrafish xenograft model of human brest cancer can be established,which demonstrates applicability for screening compounds in drug discovery studies.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2016年第1期128-132,共5页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 81202584
31400979)
山东省自主创新及成果转化专项(No 2014ZZCX02105)
山东省科技攻关项目(No 2013GHY11516)
